研究者データベース
塚越 かおりTSUKAKOSHI Kaoriツカコシ カオリ
| ■所属部署名 | 工学研究院 生命機能科学部門 | |||
| ■職名 | 助教 | |||
Last Updated :2024/09/25
業績情報
氏名・連絡先
氏名
ツカコシ カオリ, 塚越 かおり, TSUKAKOSHI Kaori
主たる所属・職名
工学研究院 生命機能科学部門, 助教
その他の所属
経歴
- -
日本学術振興会 特別研究員(DC2)
自 2011年04月, 至 2013年03月 - -
理化学研究所 脳科学総合研究センター 研究員
自 2013年04月, 至 2015年09月 - -
㈱理研バイオ―アルツハイマー病の発症前診断と予防的治療のために 研究員
自 2014年08月, 至 2015年09月 - -
東京農工大学大学院工学研究院 助教
自 2015年10月
学歴
研究分野
- A489 ナノテク・材料, A37010 生体化学
- A689 ものづくり技術（機械・電気電子・化学工学）, A27040 バイオ機能応用、バイオプロセス工学
- A189 ライフサイエンス, A49010 病態医化学
研究キーワード
科学研究費助成事業
- 学術変革領域研究(B)（総括班）
アプタマー生物学の創成による脳内恒常性維持機構の解明
自 2022年, 至 2025年 - 学術変革領域研究(B)（計画研究）
活性増強アプタマーを用いた脳内恒常性維持機構制御マウスの開発
自 2022年, 至 2025年 - 国際共同研究加速基金（国際共同研究強化(B)）
骨転移性がんの移動・浸潤におけるコラーゲン代謝の解明と治療薬創出
自 2022年, 至 2022年 - 国際共同研究加速基金（国際共同研究強化(B)）
骨転移性がんの移動・浸潤におけるコラーゲン代謝の解明と治療薬創出
自 2021年, 至 2021年 - 国際共同研究加速基金（国際共同研究強化(B)）
骨転移性がんの移動・浸潤におけるコラーゲン代謝の解明と治療薬創出
自 2020年, 至 2020年 - 若手研究
Aβオリゴマー選択的プロテアーゼの開発に基づく神経変性機構の解析
自 2019年, 至 2022年 - 国際共同研究加速基金（国際共同研究強化(B)）
骨転移性がんの移動・浸潤におけるコラーゲン代謝の解明と治療薬創出
自 2019年, 至 2019年 - 挑戦的萌芽研究
凝集・脱凝集の視点から探るアミロイドβオリゴマー生成機構
自 2015年, 至 2017年 - アプタマー技術を駆使したアルツハイマー病血液診断マーカーの探索
自 2013年, 至 2014年
論文
- Development of a DNA aptamer that binds to the complementarity-determining region of therapeutic monoclonal antibody and affinity improvement induced by pH-change for sensitive detection
Saito, Taro; Shimizu, Yutaka; Tsukakoshi, Kaori; Abe, Koichi; Lee, Jinhee; Ueno, Kinuko; Asano, Ryutaro; Jones, Brian, V; Yamada, Tomohiro; Nakano, Tatsuki; Tong, Jiaxing; Hishiki, Asami; Hara, Kodai; Hashimoto, Hiroshi; Sode, Koji; Toyo'oka, Toshimasa; Todoroki, Kenichiro; Ikebukuro, Kazunori
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
Therapeutic monoclonal antibodies (mAbs) are successful biomedicines; however, evaluation of their pharmacokinetics and pharmacodynamics demands highly specific discrimination from human immunoglobulin G naturally present in the blood. Here, we developed a novel anti-idiotype aptamer (termed A14#1) with extraordinary specificity against the anti-vascular endothelial growth factor therapeutic mAb, bevacizumab. Structural analysis of the antibody-aptamer complex showed that several bases of A14#1 recognized only the complementarity determining region (CDR) of bevacizumab, thereby contributing to its extraordinary specificity. As the CDR of bevacizumab is predicted to be highly positively charged under mildly acidic conditions and that DNA is negatively charged, the affinity of A14#1 to bevacizumab markedly increased at pH 4.7 (K-D = 44 pM) than at pH 7.4 (K-D = 12 nM). A14#1-based electrochemical detection method capable of detecting 31 pM of bevacizumab at pH 4.7 was thus developed. A14#1 could be potentially useful for therapeutic drug measurement as a novel ligand of bevacizumab.
2022年05月01日, 研究論文（学術雑誌）, 共同, 203, 0956-5663, DOI(公開)(r-map) - The state of water molecules induces changes in the topologies and interactions of G-quadruplex DNA aptamers in hydrated ionic liquid
Fujita, Kyoko; Honda, Takuya; Tsukakoshi, Kaori; Ohno, Hiroyuki; Ikebukuro, Kazunori
JOURNAL OF MOLECULAR LIQUIDS
ELSEVIER
Herein, the effects of the bound state of water molecules were investigated on the topologies and inter-actions of G-quadruplex DNA aptamers by using hydrated ionic liquids (ILs). Since intracellular water molecules are generally expected to exist in the bound state and not as free molecules that exist in the typical in vitro media, we proposed hydrated ILs as potential candidates for controlling the state of water molecules. Hydrated ILs have been reported to dissolve proteins without compromising their higher-order structures. In this study, the structures and interactions of three G-quadruplex DNA apta-mers (one thrombin-binding aptamer and two vascular endothelial growth factor 165-binding aptamers), with their target molecules were analyzed with circular dichroism (CD) spectroscopy and enzyme-linked oligonucleotide assay by changing the water content of hydrated cholinium dihydrogen phosphate (Hy [ch][dhp]). Hy[ch][dhp] allowed the effective dissolution of G-quadruplex DNA aptamers while maintain-ing their structure and binding affinity to the target molecule. The water content of Hy[ch][dhp] induced changes in the CD spectra, suggesting changes in the topology of G-quadruplex structure. Increased struc-tural stability and binding properties indicated molecular recognition and smooth dehydration progress in Hy[ch][dhp] via regulation of the state of water molecules. (c) 2022 Elsevier B.V. All rights reserved.
2022年11月15日, 研究論文（学術雑誌）, 共同, 366, 0167-7322, DOI(公開)(r-map) - Effects of G-Quadruplex Ligands on the Topology, Stability, and Immunostimulatory Properties of G-Quadruplex-Based CpG Oligodeoxynucleotides
Tu, Anh Thi Tram; Hoshi, Kazuaki; Ma, Yue; Oyama, Taiji; Suzuki, Satoko; Tsukakoshi, Kaori; Nagasawa, Kazuo; Ikebukuro, Kazunori; Yamazaki, Tomohiko
ACS CHEMICAL BIOLOGY
AMER CHEMICAL SOC
We previously reported that the formation of guanine-quadruplex (G4) structures provides phosphodiester oligodeoxynucleotides containing unmethylated cytosine-phosphate-guanine (CpG ODNs) with higher nuclease resistance and cellular uptake, thereby increasing their immunostimulation efficiency through TLR9 activation. CpG ODNs forming G4 structures (G4 CpG ODNs) are thus potential vaccine adjuvants against infectious diseases. However, the G4 structure changes topology depending on the surrounding environment. Recently, G4 ligands, which are small molecules that bind to G4 ODNs with high affinity, were reported to improve the stability of G4. In this study, we propose to increase the stability and function of G4 CpG ODNs using G4 ligands. We show the effects of two G4 ligands, named L2H2-6OTD (L2H2) and L2G2-2M2EG-6OTD (L2G2), on the topology, stability, and immunostimulatory properties of a monomeric hybrid-type G4 CpG ODN containing CpG motifs in the central loop, named GD3. We found that L2H2 helps maintain the hybrid G4 topology of GD3, whereas L2G2 induces parallel G4 formation. Both G4 ligands increase the thermodynamic and nuclease stability of GD3. However, only GD3 associated with L2H2 binds efficiently to TLR9 and evokes a higher immune response from mouse macrophage-like RAW264 cells. GD3 associated with L2G2 does not bind efficiently to TLR9 and elicits lower cytokine production. Our results demonstrate that the potential to enhance immunostimulatory properties depends on the ability of G4 ligands to maintain and stabilize the hybrid G4 of GD3. We anticipate that our findings will facilitate the development of more effective G4 CpG ODN-based vaccine adjuvants against infectious diseases.
2022年07月15日, 研究論文（学術雑誌）, 共同, 17, 7, 1554-8929, DOI(公開)(r-map), 1703, 1713 - Enhancement of DNAzymatic activity using iterative in silico maturation
Fenati, Renzo A.; Chen, Zifei; Yamagishi, Yasuko; Tsukakoshi, Kaori; Ikebukuor, Kazunori; Manian, Anjay; Russo, Salvy P.; Yamazaki, Tomohiko; Ellis, Amanda, V
JOURNAL OF MATERIALS CHEMISTRY B
ROYAL SOC CHEMISTRY
DNAzyme-based (catalytic nucleic acid) biosensing technology is recognised as a valuable biosensing tool in diagnostic medicine and seen as a cheaper, more stable alternative to antibodies or enzymes. However, like enzyme discovery, no method exists to predict DNAzyme sequences that result in high catalytic activity using computer software (in silico). In this work, iterative in silico maturation and in vitro evaluation were applied to a DNAzyme oligodeoxynucleotide (ODN) sequence to elucidate novel synthetic sequences with enhanced DNAzyme activity. An already well-known model DNAzyme, the G-quadruplex/hemin complex, was iterated over eight generations to elucidate synthetic sequences that were up to five times faster than the original parent sequence. By combining molecular dynamics simulations, we found that the POD-mimicking activities were largely affected by docking modes and the tightness of locking between complexes. Ultimately, the theoretical models showed significant sequence-dependencies.
2022年11月09日, 研究論文（学術雑誌）, 共同, 10, 43, 2050-750X, DOI(公開)(r-map), 8960, 8969 - CpG Methylation Altered the Stability and Structure of the i-Motifs Located in the CpG Islands
Oshikawa, Daiki; Inaba, Shintaro; Kitagawa, Yudai; Tsukakoshi, Kaori; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Cytosine methylation within the 5 '-C-phosphate-G-3 ' sequence of nucleotides (called CpG methylation) is a well-known epigenetic modification of genomic DNA that plays an important role in gene expression and development. CpG methylation is likely to be altered in the CpG islands. CpG islands are rich in cytosine, forming a structure called the i-motif via cytosine-cytosine hydrogen bonding. However, little is known about the effect of CpG methylation on the i-motif. In this study, The CpG methylation-induced structural changes on the i-motif was examined by thermal stability, circular dichroism (CD) spectroscopy, and native-polyacrylamide gel electrophoresis (Native-PAGE) evaluation of five i-motif-forming DNAs from four cancer-related genes (VEGF, C-KIT, BCL2, and HRAS). This research shows that CpG methylation increased the transitional pH of several i-motif-forming DNAs and their thermal stability. When examining the effect of CpG methylation on the i-motif in the presence of opposite G4-forming DNAs, CpG methylation influenced the proportion of G4 and i-motif formation. This study showed that CpG methylation altered the stability and structure of the i-motif in CpG islands.
2022年06月, 研究論文（学術雑誌）, 共同, 23, 12, DOI(公開)(r-map) - Development of Alkaline Phosphatase-Fused Mouse Prion Protein and Its Application in Toxic A beta Oligomer Detection
Tsukakoshi, Kaori; Kubo, Rikako; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Amyloid beta (A beta) oligomers play a key role in the progression of Alzheimer's disease (AD). Multiple forms of A beta assemblies have been identified by in vitro and in vivo analyses; however, it is uncertain which oligomer is highly neurotoxic. Thus, understanding the pathogenesis of AD by detecting toxic A beta oligomers is crucial. In this study, we report a fusion protein of cellular prion protein (PrPc) and alkaline phosphatase (ALP) from Escherichia coli as a sensing element for toxic A beta oligomers. Since the N-terminus domain of PrPc (residue 23-111) derived from mice is known to bind to toxic A beta oligomers in vitro, we genetically fused PrPc23-111 to ALP. The developed fusion protein, PrP-ALP, retained both the binding ability of PrPc and enzymatic activity of ALP. We showed that PrP-ALP strongly bound to high molecular weight (HMW) oligomers but showed little or no affinity toward monomers. The observation that PrP-ALP neutralized the toxic effect of A beta oligomers indicated an interaction between PrP-ALP and toxic HMW oligomers. Based on ALP activity, we succeeded in detecting A beta oligomers. PrP-ALP may serve as a powerful tool for detecting toxic A beta oligomers that may be related to AD progression.
2022年12月, 研究論文（学術雑誌）, 共同, 23, 23, DOI(公開)(r-map) - A Green Light-Regulated T7 RNA Polymerase Gene Expression System for Cyanobacteria
Shono, Chika; Ariyanti, Dwi; Abe, Koichi; Sakai, Yuta; Sakamoto, Ippei; Tsukakoshi, Kaori; Sode, Koji; Ikebukuro, Kazunori
MARINE BIOTECHNOLOGY
SPRINGER
In this study, we developed a green light-regulated T7 RNA polymerase expression system (T7 RNAP system), to provide a novel and versatile high-expression system for cyanobacteria without using any chemical inducer, realizing high expression levels comparable with previously reported for recombinant gene expression in cyanobacteria. The T7 RNAP system was constructed and introduced intoSynechocystissp. PCC6803. T7 RNAP was inserted downstream of thecpcG2promoter, which is recognized and activated by the CcaS/CcaR two-component green-light-sensing system, to compose a vector plasmid, pKT-CS01, to achieve the induction of T7 RNAP expression only under green light illumination, with repression under red light illumination. The reporter gene, superfolder green fluorescent protein (sfGFP), was inserted downstream of theT7promoter. Transcriptional analyses revealed that T7 RNAP was induced under green light but repressed under red light. Expression of thesfGFP protein derived from pKT-CS01 was observed under green light illumination and was approximately 10-fold higher than that in the control transformant, which expressedsfGFP directly under thecpcG2promoter, which is directly regulated by CcaS/CcaR, under green light illumination. Comparison with the strong promoter expression systems P(cpc560)and P(trc Delta lacO)revealed that the expression ofsfGFP by the T7 RNAP system was comparable with the levels obtained with strong promoters. These results demonstrated that the green light-regulated T7 RNAP gene expression system will be a versatile tool for future technological platform to regulate gene expression in cyanobacterial bioprocesses.
2021年02月, 研究論文（学術雑誌）, 共同, 23, 1, 1436-2228, DOI(公開)(r-map), 31, 38 - G-quadruplex: Flexible conformational changes by cations, pH, crowding and its applications to biosensing
Nishio, Maui; Tsukakoshi, Kaori; Ikebukuro, Kazunori
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
G-quadruplex (G4) is a non-canonical structure that is formed in G-rich sequences of nucleic acids. G4s play important roles in vivo, such as telomere maintenance, transcription, and DNA replication. There are three typical topologies of G4: parallel, anti-parallel, and hybrid. In general, metal cations, such as potassium and sodium, stabilize G4s through coordination in the G-quartet. While G4s have some functions in vivo, there are many reports of developed applications that use G4s. As various conformations of G4s could form from one sequence depending on varying conditions, many researchers have developed G4-based sensors. Furthermore, G4 is a great scaffold of aptamers since many aptamers folded into G4s have also been reported. However, there are some challenges about its practical use due to the difference between practical sample conditions and experimental ones. G4 conformations are dramatically altered by the surrounding conditions, such as metal cations, pH, and crowding. Many studies have been conducted to characterize G4 conformations under various conditions, not only to use G4s in practical applications but also to reveal its function in vivo. In this review, we summarize recent studies that have investigated the effects of surrounding conditions (e.g., metal cations, pH, and crowding) on G4 conformations and the application of G4s mainly in biosensor fields, and in others.
2021年04月15日, 研究論文（学術雑誌）, 共同, 178, 0956-5663, DOI(公開)(r-map) - G-quadruplex-forming aptamer enhances the peroxidase activity of myoglobin against luminol
Tsukakoshi, Kaori; Yamagishi, Yasuko; Kanazashi, Mana; Nakama, Kenta; Oshikawa, Daiki; Savory, Nasa; Matsugami, Akimasa; Hayashi, Fumiaki; Lee, Jinhee; Saito, Taiki; Sode, Koji; Khunathai, Kanjana; Kuno, Hitoshi; Ikebukuro, Kazunori
NUCLEIC ACIDS RESEARCH
OXFORD UNIV PRESS
Aptamers can control the biological functions of enzymes, thereby facilitating the development of novel biosensors. While aptamers that inhibit catalytic reactions of enzymes were found and used as signal transducers to sense target molecules in biosensors, no aptamers that amplify enzymatic activity have been identified. In this study, we report G-quadruplex (G4)-forming DNA aptamers that upregulate the peroxidase activity in myoglobin specifically for luminol. Using in vitro selection, one G4-forming aptamer that enhanced chemiluminescence from luminol by myoglobin's peroxidase activity was discovered. Through our strategy-in silico maturation, which is a genetic algorithm-aided sequence manipulation method, the enhancing activity of the aptamer was improved by introducing mutations to the aptamer sequences. The best aptamer conserved the parallel G4 property with over 300-times higher luminol chemiluminescence from peroxidase activity more than myoglobin alone at an optimal pH of 5.0. Furthermore, using hemin and hemin-binding aptamers, we demonstrated that the binding property of the G4 aptamers to heme in myoglobin might be necessary to exert the enhancing effect. Structure determination for one of the aptamers revealed a parallel-type G4 structure with propeller-like loops, which might be useful for a rational design of aptasensors utilizing the G4 aptamer-myoglobin pair.
2021年06月21日, 研究論文（学術雑誌）, 共同, 49, 11, 0305-1048, DOI(公開)(r-map), 6069, 6081 - Enhancement of the Immunostimulatory Effect of Phosphodiester CpG Oligodeoxynucleotides by an Antiparallel Guanine-Quadruplex Structural Scaffold
Safitri, Fika Ayu; Tu, Anh Thi Tram; Hoshi, Kazuaki; Shobo, Miwako; Zhao, Dandan; Witarto, Arief Budi; Sumarsono, Sony Heru; Giri-Rachman, Ernawati Arifin; Tsukakoshi, Kaori; Ikebukuro, Kazunori; Yamazaki, Tomohiko
BIOMOLECULES
MDPI
Guanine-quadruplex-based CpG oligodeoxynucleotides (G4 CpG ODNs) have been developed as potent immunostimulatory agents with reduced sensitivity to nucleases. We designed new monomeric G4 ODNs with an antiparallel topology using antiparallel type duplex/G4 ODNs as robust scaffolds, and we characterized their topology and effects on cytokine secretion. Based on circular dichroism analysis and quantification of mRNA levels of immunostimulatory cytokines, it was found that monomeric antiparallel G4 CpG ODNs containing two CpG motifs in the first functional loop, named G2.0.0, could maintain antiparallel topology and generate a high level of immunostimulatory cytokines in RAW264 mouse macrophage-like cell lines. We also found that the flanking sequence in the CpG motif altered the immunostimulatory effects. Gc2c.0.0 and Ga2c.0.0 are monomeric antiparallel G4 CpG ODNs with one cytosine in the 3 & PRIME; terminal and one cytosine/adenine in the 5 & PRIME; terminal of CpG motifs that maintained the same resistance to degradation in serum as G2.0.0 and improved interleukin-6 production in RAW264 and bone marrow-derived macrophages. The immunostimulatory activity of antiparallel G4 CpG ODNs is superior to that of linear natural CpG ODNs. These results provide insights for the rational design of highly potent CpG ODNs using antiparallel G4 as a robust scaffold.
2021年11月, 研究論文（学術雑誌）, 共同, 11, 11, DOI(公開)(r-map) - Cytotoxic A beta Protofilaments Are Generated in the Process of A beta Fibril Disaggregation
Kaku, Toshisuke; Tsukakoshi, Kaori; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Significant research on Alzheimer's disease (AD) has demonstrated that amyloid beta (A beta) oligomers are toxic molecules against neural cells. Thus, determining the generation mechanism of toxic A beta oligomers is crucial for understanding AD pathogenesis. A beta fibrils were reported to be disaggregated by treatment with small compounds, such as epigallocatechin gallate (EGCG) and dopamine (DA), and a loss of fibril shape and decrease in cytotoxicity were observed. However, the characteristics of intermediate products during the fibril disaggregation process are poorly understood. In this study, we found that cytotoxic A beta aggregates are generated during a moderate disaggregation process of A beta fibrils. A cytotoxicity assay revealed that A beta fibrils incubated with a low concentration of EGCG and DA showed higher cytotoxicity than A beta fibrils alone. Atomic force microscopy imaging and circular dichroism spectrometry showed that short and narrow protofilaments, which were highly stable in the beta-sheet structure, were abundant in these moderately disaggregated samples. These results indicate that toxic A beta protofilaments are generated during disaggregation from amyloid fibrils, suggesting that disaggregation of A beta fibrils by small compounds may be one of the possible mechanisms for the generation of toxic A beta aggregates in the brain.
2021年12月, 研究論文（学術雑誌）, 共同, 22, 23, DOI(公開)(r-map) - Detection of CpG Methylation in G-Quadruplex Forming Sequences Using G-Quadruplex Ligands
Hasegawa, Hijiri; Sasaki, Ikkei; Tsukakoshi, Kaori; Ma, Yue; Nagasawa, Kazuo; Numata, Shusuke; Inoue, Yuuki; Kim, Yeji; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Genomic DNA methylation is involved in many diseases and is expected to be a specific biomarker for even the pre-symptomatic diagnosis of many diseases. Thus, a rapid and inexpensive detection method is required for disease diagnosis. We have previously reported that cytosine methylation in G-quadruplex (G4)-forming oligonucleotides develops different G4 topologies. In this study, we developed a method for detecting CpG methylation in G4-forming oligonucleotides based on the structural differences between methylated and unmethylated G4 DNAs. The differences in G4 topologies due to CpG methylation can be discriminated by G4 ligands. We performed a binding assay between methylated or unmethylated G4 DNAs and G4 ligands. The binding abilities of fluorescent G4 ligands to BCL-2, HRAS1, HRAS2, VEGF G4-forming sequences were examined by fluorescence-based microtiter plate assay. The differences in fluorescence intensities between methylated and unmethylated G4 DNAs were statistically significant. In addition to fluorescence detection, the binding of G4 ligand to DNA was detected by chemiluminescence. A significant difference was also detected in chemiluminescence intensity between methylated and unmethylated DNA. This is the first study on the detection of CpG methylation in G4 structures, focusing on structural changes using G4 ligands.
2021年12月, 研究論文（学術雑誌）, 共同, 22, 23, DOI(公開)(r-map) - Application of a Glucose Dehydrogenase-Fused with Zinc Finger Protein to Label DNA Aptamers for the Electrochemical Detection of VEGF
Lee, Jinhee; Tatsumi, Atsuro; Tsukakoshi, Kaori; Wilson, Ellie D.; Abe, Koichi; Sode, Koji; Ikebukuro, Kazunori
SENSORS
MDPI
Aptamer-based electrochemical sensors have gained attention in the context of developing a diagnostic biomarker detection method because of their rapid response, miniaturization ability, stability, and design flexibility. In such detection systems, enzymes are often used as labels to amplify the electrochemical signal. We have focused on glucose dehydrogenase (GDH) as a labeling enzyme for electrochemical detection owing to its high enzymatic activity, availability, and well-established electrochemical principle and platform. However, it is difficult and laborious to obtain one to one labeling of a GDH-aptamer complex with conventional chemical conjugation methods. In this study, we used GDH that was genetically fused to a DNA binding protein, i.e., zinc finger protein (ZF). Fused GDH can be attached to an aptamer spontaneously and site specifically in a buffer by exploiting the sequence-specific binding ability of ZF. Using such a fusion protein, we labeled a vascular endothelial growth factor (VEGF)-binding aptamer with GDH and detected the target electrochemically. As a result, upon the addition of glucose, the GDH labeled on the aptamer generated an amperometric signal, and the current response increased dependent on the VEGF concentration. Eventually, the developed electrochemical sensor proved to detect VEGF levels as low as 105 pM, thereby successfully demonstrating the concept of using ZF-fused GDH to enzymatically label aptamers.
2020年07月, 研究論文（学術雑誌）, 共同, 20, 14, DOI(公開)(r-map) - Ethanol Detection at the Parts per Billion Level with Single-Stranded-DNA-Modified Graphene Field-Effect Transistors
Nozaki, Ryo; Ikuta, Takashi; Ueno, Kinuko; Tsukakoshi, Kaori; Ikebukuro, Kazunori; Maehashi, Kenzo
PHYSICA STATUS SOLIDI B-BASIC SOLID STATE PHYSICS
WILEY-V C H VERLAG GMBH
For realizing the early diagnosis of diseases, detection of gases in exhaled breath in parts per billion (ppb) using compact devices is necessary. Herein, single-stranded-DNA-modified graphene field-effect transistors (FETs) are fabricated and the transfer characteristics are measured. The results reveal that shifts in the transfer characteristics are obtained by introducing ethanol gas at the ppb level, indicating that single-stranded-DNA-modified graphene FETs detect the gas at the ppb level. Moreover, by modifying DNA sequence 1 or sequence 2, the positive and negative voltage shifts of transfer characteristics are observed, respectively. In contrast, the shifts are hardly obtained using DNA sequence 3, which has a rigid conformation for adsorption of molecules. These differences in the behaviors of the transfer characteristics are attributed to the change in DNA conformation when gas is adsorbed. Therefore, single-stranded-DNA-modified graphene FETs have considerable potential to detect various gases at a low concentration depending on the design of DNA sequences, enabling their promising applications in disease diagnosis.
2020年02月, 研究論文（学術雑誌）, 共同, 257, 2, 0370-1972, DOI(公開)(r-map) - G-Quadruplex Structure Improves the Immunostimulatory Effects of CpG Oligonucleotides
Hoshi, Kazuaki; Yamazaki, Tomohiko; Sugiyama, Yuuki; Tsukakoshi, Kaori; Tsugawa, Wakako; Sode, Koji; Ikebukuro, Kazunori
NUCLEIC ACID THERAPEUTICS
MARY ANN LIEBERT, INC
Single-strand oligodeoxynucleotides (ODNs) containing unmethylated cytosine-phosphate-guanine (CpG) are recognized by the toll-like receptor 9, a component of the innate immunity. Therefore, they could act as immunotherapeutic agents. Chemically modified CpG ODNs containing a phosphorothioate backbone instead of phosphodiester (PD) were developed as immunotherapeutic agents resistant to nuclease degradation. However, they cause adverse side effects, and so there is a necessity to generate novel CpG ODNs. In the present study, we designed a nuclease-resistant nonmodified CpG ODN that forms G-quadruplex structures. G-quadruplex formation in CpG ODNs increased nuclease resistance and cellular uptake. The CpG ODNs designed in this study induced interleukin-6 production in a human B lymphocyte cell line and human peripheral blood mononuclear cells. These results indicate that G-quadruplex formation can be used to increase the immunostimulatory activity of CpG ODNs having a natural PD backbone.
2019年08月01日, 研究論文（学術雑誌）, 共同, 29, 4, 2159-3337, DOI(公開)(r-map), 224, 229 - Generation of C5-desoxy analogs of tetrahydroisoquinoline alkaloids exhibiting potent DNA alkylating ability
Tanifuji, Ryo; Tsukakoshi, Kaori; Ikebukuro, Kazunori; Oikawa, Hideaki; Oguri, Hiroki
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
PERGAMON-ELSEVIER SCIENCE LTD
C5-desoxy analogs of tetrahydroisoquinoline (THIQ) alkaloids were designed and synthesized as hitherto unexplored structural variants for evaluation of their DNA alkylating activities. While chemical synthesis of the C5-desoxy analogs bearing a phenolic hydroxyl group in the A-ring of the saframycins was assumed to be laborious based on semi-synthetic modifications, a chemo-enzymatic approach allowed for concise access to the analogs. The C5-desoxy analog 7 exhibited greater DNA alkylating ability with a wider tolerance for the sequence variations compared to cyanosafracin B. The C5-desoxy A-ring having a C8 phenolic hydroxyl group, and a C1 substituent in the vicinity of the C21 aminonitrile responsible for DNA alkylation, were demonstrated to play pivotal roles in the interaction between the THIQ alkaloids and DNA.
2019年07月15日, 研究論文（学術雑誌）, 共同, 29, 14, 0960-894X, DOI(公開)(r-map), 1807, 1811 - Anti-Idiotype DNA Aptamer Affinity Purification-High-Temperature Reversed-Phase Liquid Chromatography: A Simple, Accurate, and Selective Bioanalysis of Bevacizumab
Yamada, Tomohiro; Saito, Taro; Shimizu, Yutaka; Tsukakoshi, Kaori; Hayashi, Hideki; Mizuno, Hajime; Tsuji, Daiki; Yamamoto, Keisuke; Itoh, Kunihiko; Toyo'oka, Toshimasa; Ikebukuro, Kazunori; Todoroki, Kenichiro
MOLECULES
MDPI
This study presents a simple, accurate, and selective bioanalytical method of bevacizumab detection from plasma samples based on aptamer affinity purification-high-temperature reversed-phased liquid chromatography (HT-RPLC) with fluorescence detection. Bevacizumab in plasma samples was purified using magnetic beads immobilized with an anti-idiotype DNA aptamer for bevacizumab. The purified bevacizumab was separated with HT-RPLC and detected with its native fluorescence. Using aptamer affinity beads, bevacizumab was selectively purified and detected as a single peak in the chromatogram. HT-RPLC achieved good separation for bevacizumab with a sharp peak within 10 min. The calibration curves of the two monoclonal antibodies ranged from 1 to 50 mu g/mL and showed good correlation coefficients (r(2) > 0.999). The limit of detection (LOD) and lower limit of quantification (LLOQ) values for bevacizumab were 0.15 and 0.51 mu g/mL, respectively. The proposed method was successfully applied to the bioanalysis of the plasma samples obtained from the patients with lung cancer and may be extended to plan optimal therapeutic programs and for the evaluation of biological equivalencies in the development of biosimilars.
2019年03月01日, 研究論文（学術雑誌）, 共同, 24, 5, 1420-3049, DOI(公開)(r-map) - High-Throughput Bioanalysis of Bevacizumab in Human Plasma Based on Enzyme-Linked Aptamer Assay Using Anti-Idiotype DNA Aptamer
Yamada, Tomohiro; Saito, Taro; Hill, Yoshia; Shimizu, Yutaka; Tsukakoshi, Kaori; Mizuno, Hajime; Hayashi, Hideki; Ikebukuro, Kazunori; Toyo'oka, Toshimasa; Todoroki, Kenichiro
ANALYTICAL CHEMISTRY
AMER CHEMICAL SOC
We propose a highly selective, sensitive, accurate, and high throughput bioanalysis method for bevacizumab utilizing an anti-idiotype DNA aptamer. With this method, bevacizumab in a plasma sample was reacted in a 96-well plate immobilized with the aptamer and further reacted with a protein A HRP conjugate. The resulting HRP activity was colorimetrically detected using a microplate reader. The calibration curve of bevacizumab ranged from 0.05 to 5.0 mu g/mL, and showed a good correlation coefficient (r(2) = 1.000). The limit of detection was 2.09 ng/mL. We also demonstrated both the possibility of highly sensitive detection using luminol chemiluminescence and the repeated use of affinity plates. The proposed method is applicable for planning optimal therapeutic programs and for an evaluation of the biological equivalencies in the development of biosimilars.
2019年02月19日, 研究論文（学術雑誌）, 共同, 91, 4, 0003-2700, DOI(公開)(r-map), 3125, 3130 - Selection and Characterization of DNA Aptamers Against FokI Nuclease Domain.
Nishio M, Yamagishi A, Tsukakoshi K, Kato Y, Nakamura C, Ikebukuro K.
Methods Mol Biol.
2018年, 研究論文（学術雑誌）, 共同, DOI(公開)(r-map) - Riboregulator elements as tools to engineer gene expression in cyanobacteria
Kinuko Ueno, Kaori Tsukakoshi, Kazunori Ikebukuro
Applied Microbiology and Biotechnology
2018年09月, 研究論文（学術雑誌）, 共同, 102, 18, DOI(公開)(r-map), 7717, 7723 - Selection and Characterization of DNA Aptamers Against FokI Nuclease Domain
Maui Nishio, Ayana Yamagishi, Kaori Tsukakoshi, Yoshio Kato, Chikashi Nakamura, Kazunori Ikebukuro
Methods in Molecular Biology-Zinc Finger Proteins
2018年08月, 研究論文（学術雑誌）, 共同, DOI(公開)(r-map), 165, 174 - Esterification of PQQ Enhances Blood-Brain Barrier Permeability and Inhibitory Activity against Amyloidogenic Protein Fibril Formation
Kaori Tsukakoshi, Wataru Yoshida, Masaki Kobayashi, Natsuki Kobayashi, Jihoon Kim, Toshisuke Kaku, Toshitsugu Iguchi, Kazuo Nagasawa, Ryutaro Asano, Kazunori Ikebukuro, Koji Sode
ACS Chemical Neuroscience
2018年08月, 研究論文（学術雑誌）, 共同, DOI(公開)(r-map) - CpG Methylation Changes G-Quadruplex Structures Derived from Gene Promoters and Interaction with VEGF and SP1
Kaori Tsukakoshi, Shiori Saito, Wataru Yoshida, Shinichi Goto, Kazunori Ikebukuro
Molecules
2018年04月, 研究論文（学術雑誌）, 共同, 23, 4, DOI(公開)(r-map), 944 - Improving the induction fold of riboregulators for cyanobacteria.
Ippei Sakamoto, Koichi Abe, Sumiya Kawai, Kaori Tsukakoshi, Yuta Sakai, Koji Sode, Kazunori Ikebukuro
RNA Biology
2018年01月, 研究論文（学術雑誌）, 共同, 5, DOI(公開)(r-map), 1, 6 - Development of HGF-binding aptamers with the combination of G4 promoter-derived aptamer selection and in silico maturation
Tomomi Yokoyama, Kaori Tsukakoshi, Wataru Yoshida, Taiki Saito, Kentaro Teramoto, Nasa Savory, Koichi Abe, Kazunori Ikebukuro
Biotechnology and Bioengineering
WILEY
We describe the selection of aptamers based on bioinformatics-based approaches without Systematic Evolution of Ligands by EXponential enrichment (SELEX). SELEX is a potent method; however, it is time intensive and the PCR-amplification step, which is essential step for SELEX, leads to the loss of good aptamers. We have developed an aptamer-screening method, G4 promoter-derived aptamer selection (G4PAS), and an aptamer-improving method, in silico maturation (ISM). They are based on in silico sequence selection and computer assisted directed evolution, respectively. In this study, we succeeded in identifying new aptamers against hepatocyte growth factor (HGF) by G4PAS as well as improving the specificity of the HGF aptamers by ISM. Using ISM improved the specificity of the aptamer for HGF by up to 45-fold in comparison with the original aptamer. These methods enable easy and efficient identification of good aptamers, and the combination of G4PAS with ISM can thus serve as a potent approach for aptamer identification. Biotechnol. Bioeng. 2017;114: 2196-2203. (c) 2017 Wiley Periodicals, Inc.
2017年10月, 研究論文（学術雑誌）, 共同, 114, 10, 0006-3592, DOI(公開)(r-map), 2196, 2203 - Applying a riboregulator as a new chromosomal gene regulation tool for higher glycogen production in Synechocystis sp. PCC 6803
Kinuko Ueno, Yuta Sakai, Chika Shono, Ippei Sakamoto, Kaori Tsukakoshi, Yukako Hihara, Koji Sode, Kazunori Ikebukuro
Applied Microbiology and Biotechnology
2017年10月, 研究論文（学術雑誌）, 共同, 101, 23-24, DOI(公開)(r-map), 8465, 8474 - Development of aptamers against unpurified proteins
Shinichi Goto, Kaori Tsukakoshi, Kazunori Ikebukuro
Biotechnology and Bioengineering
2017年08月, 研究論文（学術雑誌）, 共同, 114, 12, DOI(公開)(r-map), 2706, 2716 - DNA aptamers against FokI nuclease domain for genome editing applications
Maui Nishio, Daisuke Matsumoto, Yoshio Kato, Koichi Abe, Jinhee Lee, Kaori Tsukakoshi,Ayana Yamagishi, Chikashi Nakamura, Kazunori Ikebukuro
Biosensors and Bioelectronics
ELSEVIER ADVANCED TECHNOLOGY
Genome editing with site-specific nucleases (SSNs) can modify only the target gene and may be effective for gene therapy. The main limitation of genome editing for clinical use is off-target effects; excess SSN5 in the cells and their longevity can contribute to off-target effects. Therefore, a controlled delivery system for SSN5 is necessary. Fold nuclease domain (FokI) is a common DNA cleavage domain in zinc finger nuclease (ZFN) and transcription activator-like effector nuclease. Previously, we reported a zinc finger protein delivery system that combined aptamer-fused, double-strand oligonucleotides and nanoneedles. Here, we report the development of DNA aptamers that bind to the target molecules, with high affinity and specificity to the FokI. DNA aptamers were selected in six rounds of systematic evolution of ligands by exponential enrichment. Aptamers F6#8 and #71, which showed high binding affinity to Fokl (K-d=82 nM, 74 nM each), showed resistance to nuclease activity itself and did not inhibit nuclease activity. We immobilized the ZFN-fused GFP to nanoneedles through these aptamers and inserted the nanoneedles into HEK293 cells. We observed the release of ZFN-fused GFP from the nanoneedles in the presence of cells. Therefore, these aptamers are useful for genome editing applications such as controlled delivery of SSNs.
2017年07月, 研究論文（国際会議プロシーディングス）, 共同, 93, 0956-5663, DOI(公開)(r-map), 26, 31 - Sensitive and Homogeneous Detection System with Aptamer-Based Biosensor
Kaori Tsukakoshi, Kazunori Ikebukuro
Sensors and Materials
2016年10月, 研究論文（学術雑誌）, 共同, 28, 10, DOI(公開)(r-map), 1083, 1089 - Structural regulation by a G-quadruplex ligand increases binding abilities of G-quadruplex-forming aptamers
Kaori Tsukakoshi, Yuri Ikuta, Koichi Abe, Wataru Yoshida, Keisuke Iida, Yue Ma, Kazuo Nagasawa, Koji Sode, Kazunori Ikebukuro
Chemical Communications
2016年10月, 研究論文（学術雑誌）, 共同, 52, 85, DOI(公開)(r-map), 12646, 12649 - Improvement of the VEGF binding ability of DNA aptamers through in silico maturation and multimerization strategy
Takahiro Fukaya, Koichi Abe, Nasa Savory, Kaori Tsukakoshi, Wataru, Stefano Ferri, Koji Sode, Kazunori Ikebukuro
Journal of Biotechnology
2015年10月, 研究論文（学術雑誌）, 共同, 212, DOI(公開)(r-map), 99, 105 - Simultaneous Improvement of Specificity and Affinity of Aptamers Against Streptococcus mutans by In Silico Maturation for Biosensor Development
Nasa Savory, Yayoi Takahashi, Kaori Tsukakoshi, Hijiri Hasegawa, Madoka Takase, Koichi Abe, Wataru Yoshida, Stefano Ferri, Shizuko Kumazawa, Koji Sode, Kazunori Ikebukuro
Biotechnology and Bioengineering
2014年03月, 研究論文（学術雑誌）, 共同, 111, 3, DOI(公開)(r-map), 454, 461 - アミロイドβ代謝の分子機構から紐解くアルツハイマー病
塚越かおり, 西道隆臣
生体の科学
2014年02月, 共同, 65, 1, 69, 73 - In Silico Maturation of Binding-Specificity of DNA Aptamers Against Proteus mirabilis
Nasa Savory, Danielle Lednor, Kaori Tsukakoshi, Koichi Abe, Wataru Yoshida, Stefano Ferri, Brian V Jones, Kazunori Ikebukuro
Biotechnology and Bioengineering
2013年10月, 研究論文（学術雑誌）, 共同, 110, 10, DOI(公開)(r-map), 2573, 2580 - アミロイド形成蛋白質に結合する核酸リガンド、アプタマー開発の現状：アミロイドオリゴマー結合アプタマーの同定
塚越かおり，阿部公一，早出広司，池袋一典
Dementia Japan
2012年09月, 共同, 26, 325, 333 - Selection of DNA Aptamers That Recognize alpha-Synuclein Oligomers Using a Competitive Screening Method
Kaori Tsukakoshi, Koichi Abe, Koji Sode, Kazunori Ikebukuro
Analytical Chemistry
2012年07月, 研究論文（学術雑誌）, 共同, 84, 13, DOI(公開)(r-map), 5542, 5547 - Non-label homogeneous protein detection based on laser interferometric photo-thermal displacement measurement using aptamers
Kaori Tsukakoshi, Daisuke Ogasawara, Eiji Takahashi, Ryo Katayama, Kazunori Ikebukuro
Biotechnology Journal
2011年01月, 研究論文（学術雑誌）, 共同, 6, 1, DOI(公開)(r-map), 101, 106 - Screening of DNA aptamer which binds to alpha-synuclein
Kaori Tsukakoshi, Ryuichi Harada, Koji Sode, Kazunori Ikebukuro
Biotechnology Letters
2010年01月, 研究論文（学術雑誌）, 共同, 32, 5, DOI(公開)(r-map), 643, 648 - Development of a liquid chromatography-based versatile bioanalysis for bevacizumab based on pretreatment combining aptamer affinity purification and centrifugal ultrafiltration concentration
Todoroki, Kenichiro; Hamada, Daichi; Yamada, Tomohiro; Saito, Taro; Shimizu, Yutaka; Sugiyama, Eiji; Mizuno, Hajime; Hayashi, Hideki; Tsukakoshi, Kaori; Ikebukuro, Kazunori
ANALYTICAL SCIENCES
SPRINGERNATURE
We report on the development of a versatile and accurate bioanalytical method for bevacizumab using a pretreatment method combining affinity purification with anti-idiotypic DNA aptamers and centrifugal ultrafiltration concentration, followed by liquid chromatography (LC)-fluorescence analysis. An affinity purification method using Sepharose beads as an affinity support removed immunoglobulin G and a large amount of coexisting substances in the serum sample. Purified bevacizumab was separated as a single peak by conventional LC and detected fluorometrically, showing good linearity (R2 = 0.999) in the range of 5-200 & mu;g/mL, sufficient to analyze bevacizumab concentrations in the blood of bevacizumab-treated patients. By combining this purification method with a concentration method using a centrifugal filtration device that inhibits non-specific adsorption of bevacizumab, the quantitative range was reduced by a factor of 10 while showing good linearity (R2 = 0.999) in the 0.5-20 & mu;g/mL range. The developed analytical method is expected to be used not only for general bioanalysis of therapeutic mAbs in clinical settings, but also for next-generation antibody drugs that show drug efficacy at low concentrations and for analysis of trace samples.
2023年11月, 研究論文（学術雑誌）, 共同, 39, 11, 0910-6340, DOI(公開)(r-map), 1805, 1811 - Real-time monitoring of the amyloid β1-42 monomer-to-oligomer channel transition using a lipid bilayer system
Numaguchi, Yuri; Tsukakoshi, Kaori; Takeuchi, Nanami; Suzuki, Yuki; Ikebukuro, Kazunori; Kawano, Ryuji; Wand, Josh
PNAS NEXUS
OXFORD UNIV PRESS
This study describes the observation of the transformation of monomeric amyloid β1–42 (Aβ42) into oligomers in a lipid membrane utilizing a lipid bilayer system for electrophysiological measurement. The relevance of oligomers and protofibrils in Alzheimer's disease (AD) is underscored given their significant neurotoxicity. By closely monitoring the shift of Aβ42 from its monomeric state to forming oligomeric channels in phospholipid membranes, we noted that this transformation transpired within a 2-h frame. We manipulated the lipid membrane's constitution with components such as glycerophospholipid, porcine brain total lipid extract, sphingomyelin (SM), and cholesterol (Chol.) to effectively imitate nerve cell membranes. Interesting findings showcased Chol.'s ability to foster stable oligomeric channel formation in the lipid membrane, with SM and GM1 lipids potentially enhancing channel formation as well. Additionally, the study identified the potential of a catechin derivative, epigallocatechin gallate (EGCG), in obstructing oligomerization. With EGCG present in the outer solution of the Aβ42-infused membrane, a noteworthy reduction in channel current was observed, suggesting the successful inhibition of oligomerization. This conclusion held true in both, prior and subsequent, stages of oligomerization. Our findings shed light on the toxicity of oligomers, promising invaluable information for future advancements in AD treatment strategies.
2023年12月14日, 研究論文（学術雑誌）, 共同, 3, 1, DOI(公開)(r-map) - Identification of novel amyloidosis in dogs: α-S1-casein acquires amyloidogenicity in mammary tumor by overexpression and N-terminal truncation
Murakami, Tomoaki; Kaku, Toshisuke; Tsukakoshi, Kaori; Iwaide, Susumu; Itoh, Yoshiyuki; Hisada, Miki; Nomura, Kohji; Kubo, Rikako; Ikebukuro, Kazunori; Sassa-O'Brien, Yukiko; Kametani, Fuyuki
VETERINARY PATHOLOGY
SAGE PUBLICATIONS INC
Mammary tumor-associated amyloidosis (MTAA) in dogs is characterized by amyloid deposition in the stroma of mammary adenoma or carcinoma; however, the amyloid precursor protein remains unknown. We attempted to identify an amyloid precursor protein and elucidated its etiology by characterizing 5 cases of canine MTAA. Proteomic analyses of amyloid extracts from formalin-fixed paraffin-embedded specimens revealed alpha-S1-casein (CASA1) as a prime candidate and showed the N-terminal truncation of canine CASA1. Both immunohistochemistry and immunoelectron microscopy showed that amyloid deposits or fibrils in MTAA cases were positive for CASA1. Reverse transcription-polymerase chain reaction and quantitative polymerase chain reaction revealed the complete mRNA sequence encoding CASA1, whose expression was significantly higher in the amyloid-positive group. The recombinant protein of the N-terminal-truncated canine CASA1 and the synthetic peptides derived from canine and human CASA1 formed amyloid-like fibrils in vitro. Structural prediction suggested that the N-terminal region of CASA1 was disordered. Previously, full-length CASA1 was reported to inhibit the amyloidogenesis of other proteins; however, we demonstrated that CASA1 acquires amyloidogenicity via excessive synthesis followed by truncation of its disordered N-terminal region. By identifying a novel in vivo amyloidogenic protein in animals and revealing key mechanistic details of its associated pathology, this study provides valuable insights into the integrated understanding of related proteopathies.
2023年03月, 研究論文（学術雑誌）, 共同, 60, 2, 0300-9858, DOI(公開)(r-map), 203, 213
著書
- Advances in Synthetic Biology.
K. Ueno, K. Tsukakoshi, and K. Ikebukuro.
Engineering of Riboregulators for Gene Regulation as a Tool for Synthetic Biology
Springer Singapore
In Advances in Synthetic Biology pp. 173,
2020年04月13日, 978-981-15-0081-7
研究発表、招待講演等
- Virus detection and on-site visualization of infected spot by sequential enzymatic reaction between antibody-enzyme complex and aptamer-DNAzyme
ISNAC2023
2023年11月, 口頭発表（一般） - Exploration of DNA binding proteins for assembling covalently bound DNA-protein complexes and the application for biosensing system
ISNAC 2023
2023年11月, 口頭発表（一般） - A Biosensor Platform for On-site Virus Visualization
244th ECS Meeting
2023年10月, 口頭発表（一般） - Exploration of DNA binding proteins as a versatile tool for fabrication of DNA-protein complexes and its application to biosensing system
244th ECS Meeting
2023年10月, 口頭発表（一般） - Improvement of G-quadruplex-forming DNA aptamer for α-synuclein oligomer by loop modification
ISNAC 2023
2023年11月, ポスター発表 - Detection of CpG methylation based on structural change of G-quadruplex forming DNA oligonucleotide and its binding to heme proteins
ISNAC 2023
2023年11月, ポスター発表 - The methylated DNA changes the rate of strand displacement DNA polymerase amplification.
ISNAC 2023
2023年11月, ポスター発表 - ファブリー病のバイオマーカーであるマルベリー小体の検出系構築
第46回日本分子生物学会年会
2023年12月, ポスター発表 - SARS CoV-2 ゲノムRNAにおけるグアニン四重鎖構造が、転写と翻訳に与える影響の評価系の構築
第46回日本分子生物学会年会
2023年12月, ポスター発表 - Development of ligands that specifically bind to oligomeric amyloid protein
IPR-iCeMS joint seminar
2023年08月28日, 口頭発表（招待・特別） - 酵素活性を制御するDNAアプタマーの開発
第96回日本生化学会大会
2S07a「ケミカルバイオロジーが挑む生体分子の化学修飾」
2023年11月01日, 口頭発表（招待・特別） - アプタマー生物学を活用した脳内タンパク質の機能制御
第13回 CSJ化学フェスタ2023
文科省科研費学術変革領域研究（B）「多元応答ゲノム」「アプタマー生物学」「革新ラマン」特別企画：細胞内の生命分子の機能を知り、操り、視る技術を開発する
2023年10月17日, シンポジウム・ワークショップ パネル（公募） - タンパク質機能を制御するDNAアプタマーの開発：「アプタマー生物学」の創成へ
第41回日本認知症学会学術集会／第37回日本老年精神医学会
2022年11月27日, シンポジウム・ワークショップ パネル（公募） - Proximity ligation assay using amyloid oligomer-binding aptamer with antibody indicates Aβ1-42 and Aβ3pE-42 oligomer formation in brain
ISNAC2022
2022年11月02日, ポスター発表 - Aptameric enzyme subunit enhances the peroxidase activity of myoglobin against luminol
FIBER日本核酸化学会若手フォーラム
2021年08月, その他 - Proximity ligation assay combining aptamer with antibody for detection of amyloid-beta oligomers in brains
ISNAC2021
2021年11月, ポスター発表 - Specific detection of a therapeutic antibody using an anti-idiotype DNA aptamer and drastic improvement of its affinity by pH optimization
BIOSENSORS 2021
2021年07月, ポスター発表 - Analysis of inosine introduced aptamer against SARS-CoV-2 S1 protein RBD
第15回バイオ関連化学シンポジウム
2021年09月, ポスター発表 - Evaluation of the structural effects of CpG methylation on the dopamine receptor gene(DRD2)
2021年電気化学秋季大会
2021年09月, 口頭発表（一般） - Evaluation of the aptamer binding to the target molecule in the mist using quartz crystal units for detection of virus in the air
2021年電気化学秋季大会
2021年09月, 口頭発表（一般） - グアニン四重鎖構造を用いたDNAアジュバントの開発
日本核酸医薬学会第6回年会
2021年06月, ポスター発表 - Development of functional oligonucleotides forming G-quadruplex structure by using topological structure evaluation based on CD spectrum analysis combined with principal component analysis
日本核酸医薬学会第6回年会
2021年06月, ポスター発表 - チャネル電流計測法を用いた脂質膜上におけるAβのポア構造形成過程の解析
第３９回日本認知症学会学術集会
2020年11月, ポスター発表 - 病態生理学的知見に基づいた毒性アミロイドβオリゴマー識別リガンドの開発
第14回バイオ関連化学シンポジウム
2020年09月, 口頭発表（一般） - イヌの乳腺腫瘍のアミロイド沈着より見出だされたα-S1-カゼインの凝集特性評価
日本化学会第100春季年会
2020年03月, 口頭発表（一般） - Improvement of fluorescent aptamer focusing on its G-quadurplex and design of pH-sensitive Baby Spinach aptamer
日本化学会第100春季年会
2020年03月, 口頭発表（一般） - アプタマーを用いたAβオリゴマーの組織染色法の検討
第３８回日本認知症学会学術集会
2019年11月, ポスター発表 - Aβオリゴマーの検出に向けたアルカリホスファターゼ融合プリオンタンパク質の開発
第３８回日本認知症学会学術集会
2019年11月, ポスター発表 - Aβ線維の脱凝集によるAβオリゴマー形成の検討
第３８回日本認知症学会学術集会
2019年11月, ポスター発表 - Gas sensing based on DNA-modified graphene devices
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Chemo-enzymatic rapid synthesis of tetrahydroisoquinoline alkaloids exhibiting reversible DNA alkylating ability
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Evaluation of the effect of CpG methylation on i-motif structure located in the CpG islands
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Topological structure evaluation of G-quadruplexes using high-throughput CD system
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Enhancement of peroxidase activity of myoglobin by parallel G-quadruplex forming aptamer
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Effects of cation and G-quadruplex(G4) ligands on topology and binding ability of Cas9-binding aptamer
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Improvement in binding property of the anti-idiotype aptamer against bevacizumab based on structural information
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Improvement and design of pH-sensitive Baby Spinach aptamer by fusing triplex forming sequence
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, ポスター発表 - Ligand-based functional improvement of G-quadruplex- forming DNA aptamers
The 46th International Symposium on Nucleic Acids Chemistry
2019年10月, 口頭発表（一般） - 蛍光発光アプタマーに対する三重鎖形成配列の付加による蛍光強度の増強
第71回日本生物工学会大会
2019年09月, 口頭発表（一般） - プロモーター領域に存在する i-motif の安定性に CpG のメチル化が与える効果
第13回バイオ関連シンポジウム
2019年09月, 口頭発表（一般） - DNAアプタマーを用いたBevacizumabの検出法の開発
2019年電気化学秋季大会
2019年09月, 口頭発表（一般） - ミオグロビン結合アプタマーによる、ミオグロビンのペルオキシダーゼ活性の増強
2019年電気化学秋季大会
2019年09月, 口頭発表（一般） - イヌの乳腺腫瘍随伴アミロイドーシスにおける前駆タンパク同定の試み(続報)
第七回日本アミロイドーシス学会学術集会
2019年08月, ポスター発表 - 円二色性スペクトル解析に基づくαシヌクレインオリゴマー結合アプタマーの構造活性評価
日本核酸医薬学会第５回年会
2019年07月, ポスター発表 - PROXIMITY LIGATION ASSAY TO SPECIFICALLY DETECT AMYLOID BETA OLIGOMERS
14th International Conference on Alzheimer’s and Parkinson’s Diseases (AD/PD 2019)
2019年03月, ポスター発表 - DEVELOPMENT OF ALKALINE PHOSPHATASE-FUSED PRION PROTEIN FOR SPECIFIC DETECTION OF AMYLOID BETA OLIGOMERS
14th International Conference on Alzheimer’s and Parkinson’s Diseases (AD/PD 2019)
2019年03月, ポスター発表 - Detection of amyloidogenic protein oligomers with combination of DNA aptamers and antibodies
FNA Perth 2018
2018年11月, 口頭発表（招待・特別） - Screening of DNA aptamers against synthetic lipopeptide UPM-1 for Ureaplasma detection
The 45th International Symposium on Nucleic Acids Chemistry
2018年11月, ポスター発表 - Screening of anti-idiotype aptamer against Nivolumab
The 45th International Symposium on Nucleic Acids Chemistry
2018年11月, ポスター発表 - Mutational analysis and improvement of Baby Spinach focusing on its G-quadruplex structure
The 45th International Symposium on Nucleic Acids Chemistry
2018年11月, 口頭発表（一般） - トロンビン-アンチトロンビン複合体の検出を目指した多価化アプタマーの設計とG-quadruplex結合リガンドを用いた結合能の向上
第41回分子生物学会年会
2018年11月, ポスター発表 - アプタマー修飾磁気ビーズを用いた αシヌクレインオリゴマー検出法の開発
第37回日本認知症学会学術集会
2018年10月, ポスター発表 - Proximity Ligation AssayによるAβ1-42オリゴマーの検出
第37回日本認知症学会学術集会
2018年10月, ポスター発表 - Bevacizumabに対する抗いディオタイプアプタマーの結合特性の改良
第70回日本生物工学会大会
2018年09月, 口頭発表（一般） - 蛍光発光アプタマーSpinachに対する変異導入とその蛍光への影響評価
第70回日本生物工学会大会
2018年09月, 口頭発表（一般） - Proximity ligation assayによるアミロイドβオリゴマーの検出
第12回バイオ関連化学シンポジウム
2018年09月, 口頭発表（一般） - アルカリホスファターゼ融合プリオンタンパク質質を用いたアミロイドβオリゴマー検出法の開発
第12回バイオ関連化学シンポジウム
2018年09月, ポスター発表 - トロンビン-アンチトロンビン複合体に結合するアプタマーの設計とG-quadruplex結合リガンドを用いた結合能の向上
第12回バイオ関連化学シンポジウム
2018年09月, ポスター発表 - Smart aptamers forming G-quadruplex: Structural and functional change of aptamers forming G-quadruplex in response to surrounding conditions and its regulation with the ligands
ACS national meeting
2018年08月, 口頭発表（一般） - Primary and tertiary structure specific detection of amyloid β oligomers by proximity ligation assay
Biosensors 2018
2018年06月, ポスター発表 - Glucose dehydrogenase-fused zinc finger protein to label DNA aptamers for electrochemical detection
Biosensors 2018
2018年06月, 口頭発表（一般） - Applying riboregulator to knock down chromosomal gene cyabrB2 in Synechocystis sp.PCC 6803 for higher glycogen production.
RNA conference 2018
2018年05月, ポスター発表 - Structural regulation of G-quadruplex-forming aptamers by a G-quadruplex ligand to control its binding ability
日本化学会第98春季大会
2018年03月, 口頭発表（一般） - ミオグロビン結合アプタマーによる、ミオグロビンのペルオキシダーゼ活性の増強
日本化学会第98春季大会
2018年03月, 口頭発表（一般） - トロンビン-アンチトロンビン結合アプタマーのG-quadruplex構造と結合能との相関解析
電気化学会第85回大会
2018年03月, 口頭発表（一般） - シアノバクテリアSynechocystis sp. PCC 6803におけるリボレギュレーターを用いたcyabrB2遺伝子の発現制御およびグリコーゲン高生産株の構築
藍藻の分子生物学2017
2017年12月, ポスター発表 - Aβオリゴマーの高感度検出に向けたProximity Ligation Assayの開発
第３６回日本認知症学会学術集会
2017年11月, ポスター発表 - The effects of CpG methylation on the structure and the binding property against proteins of G-quadruplex forming DNAs
The 44th International Symposium on Nucleic Acids Chemistry
2017年09月, 口頭発表（一般） - Smart aptamers changing its structure and binding a nity to the target protein responding to cations
The 44th International Symposium on Nucleic Acids Chemistry
2017年09月, ポスター発表 - Mutational analysis of Baby Spinach focusing on its G-quadruplex structure
The 44th International Symposium on Nucleic Acids Chemistry
2017年09月, ポスター発表 - Competitive binding of thrombin aptamers and antithrombin against thrombin.
The 44th International Symposium on Nucleic Acids Chemistry
2017年09月, ポスター発表 - Targer DNA detection using alkaline phosphatase fused zinc finger protein for diagnosis
The 44th International Symposium on Nucleic Acids Chemistry
2017年09月, ポスター発表 - リボレギュレーターを用いたcyabrB2遺伝子の発現制御によるグリコーゲン高生産シアノバクテリアの構築
第69回生物工学会大会
2017年09月, 口頭発表（一般） - G-quadruplex構造及びタンパク質の相互作用に対するDNAメチル化の影響解析
第69回生物工学会大会
2017年09月, ポスター発表 - アバスチンに対する抗イディオタイプアプタマーの結合の特性評価
第11回バイオ関連化学シンポジウム
2017年09月, ポスター発表 - Proximity Ligation Assayによるアミロイドβ特異的検出
第11回バイオ関連化学シンポジウム
2017年09月, ポスター発表 - アミロイドβオリゴマーの検出に向けたアルカリホスファターゼ融合プリオンタンパク質の開発
第11回バイオ関連化学シンポジウム
2017年09月, ポスター発表 - アルカリフォスファターゼ融合ジンクフィンガー蛋白質の開発と標的DNAの検出
第11回バイオ関連化学シンポジウム
2017年09月, ポスター発表 - アプタマーを用いたトロンビン・アンチトロンビン複合体の検出
電気化学秋季大会
2017年09月, 口頭発表（一般） - Development of alkaline phosphatase fused zinc finger protein for target DNA detection
BES-2017 (XXIV International Symposium on Bioelectrochemistry and Bioenergetics)
2017年07月, 口頭発表（一般） - Applying riboregulator to knock down chromosomal gene cyabrB2 in Synechocystis sp.PCC 6803 for higher glycogen production
11th Asia-Pacific Marine Biotechnology Conference (APMBC) 2017
2017年05月, 口頭発表（一般） - 標的DNA検出に向けたアルカリフォスファターゼ融合ジンクフィンガー蛋白質の開発
日本化学会第97春季年会
2017年03月, 口頭発表（一般） - アミロイドオリゴマー結合アプタマーを用いた、核酸増幅に基づく水溶性アミロイドβオリゴマーの検出
日本化学会第97春季年会
2017年03月, 口頭発表（一般） - 非天然塩基の導入によるトロンビン結合アプタマーの結合能の改良
日本化学会第97春季年会
2017年03月, 口頭発表（一般） - Development of the electrochemical detection system of thrombin activity
PepCon-2017
2017年03月, ポスター発表 - BINDING PROPERTIES OF AMYLOID OLIGOMER-BINDING DNA APTAMERS AGAINST AMYLOID BETA OLIGOMERS
13th International Conference on Alzheimer’s and Parkinson’s Diseases (AD/PD 2017)
2017年03月, ポスター発表 - シトシンメチル化がG-quadruplexとタンパク質の結合に及ぼす影響の評価
第39回日本分子生物学会年会
2016年11月, ポスター発表 - 抗体医薬品を特異的に認識する抗イディオタイプアプタマーの探索とその特性評価
第39回日本分子生物学会年会
2016年11月, ポスター発表 - Development of the Electrochemical Detection System Using the Combination of Aptamer and Enzyme
PRiME 2016
2016年10月, 口頭発表（一般） - タンパク質とG-quadruplexとの結合にシトシンのメチル化が与える影響の解析
第10回バイオ関連化学シンポジウム
2016年09月, ポスター発表 - リボレギュレーターを用いた内在性遺伝子cyabrB2 の転写および発現制御
第10回バイオ関連化学シンポジウム
2016年09月, ポスター発表 - 高感度検出への応用を目指したアルカリホスファターゼ阻害アプタマーの探索
第10回バイオ関連化学シンポジウム
2016年09月, ポスター発表 - グアニン四重鎖特異的リガンドを用いたDNAアプタマーの構造制御
第10回バイオ関連化学シンポジウム
2016年09月, 口頭発表（一般） - Construction of photo regulation system of protein expression in Synechocystis sp. PCC 6803
The 43rd International Symposium on Nucleic Acids Chemistry
2016年09月, ポスター発表 - Effect of G-quadruplex ligand on the topology of G-quadruplex forming aptamer and its affinity to the target molecules
The 43rd International Symposium on Nucleic Acids Chemistry
2016年09月, ポスター発表 - Screening and characterization of aptamer for myoglobin
The 43rd International Symposium on Nucleic Acids Chemistry
2016年09月, ポスター発表 - Improvement of binding affinity of G-quadruplex forming aptamers
Biosensors 2016
2016年05月, ポスター発表 - Interaction between G-quadruplex (G4)-forming aptamer and heme protein
Biosensors 2016
2016年05月, ポスター発表 - Development of multivalent aptamers for high-sensitive detection of target proteins
Biosensors 2016
2016年05月, ポスター発表 - アミロイドオリゴマーを認識する核酸リガンドDNAアプタマーの開発
第30回日本認知症学会学術集会
2011年11月, 口頭発表（招待・特別）
外部研究資金等
- アルツハイマー病の迅速診断に向けた?液バイオマーカー・マルチセンシングバイオセンサーの開発
受託研究, 自 2023年01月24日, 至 2023年03月31日 - 奨学寄附金
奨学寄附金, 自 2022年, 至 2022年 - 奨学寄附金
奨学寄附金, 自 2018年, 至 2018年 - 奨学寄附金
奨学寄附金, 自 2018年, 至 2018年 - 奨学寄附金
奨学寄附金, 自 2017年, 至 2017年 - 奨学寄附金
奨学寄附金, 自 2016年, 至 2016年
メディア報道
- Researchers uncover novel amyloidosis α-S1-casein forms amyloid by overexpression and N-terminal truncation in canine mammary tumor
A collaboration led by scientists at Tokyo University of Agriculture and Technology (TUAT), Japan, has discovered a novel amyloid protein from canine mammary tumors.
EurekAlert!／Phys.Org／News-Medical.Net
自 2023年01月23日, 至 2023年01月24日 - Alzheimer's early detection through biomarkers - award for Swansea University and Tokyo University project
This new major project will see Dr Sharma lead a consortium of leading scientists from Swansea University, Imperial College London and The University of Glasgow. Dr Kaori Tsukakoshi will steer researchers from Japan with Tokyo University of Agriculture & Technology and National Institutes for Quantum Science & Technology. Together they will create a Point-of-Care Testing Kit to facilitate early diagnosis and monitoring of disease progression in primary clinics or home settings.
EurekAlert!／NEWS MEDICAL LIFE SCIENCE／MEEFRO
自 2023年03月23日, 至 2023年03月27日 - 運転中の心筋梗塞、予兆を早期検知…デンソーなどDNAの機能を発見
デンソーと東京農工大学、理化学研究所、ノースカロライナ大学は、特殊な構造（グアニン四重鎖構造）を持つDNAがミオグロビンタンパク質の持つ酵素活性を増強する機能があることを発見したことが紹介される。
yahooニュース／Response／msnニュース
自 2021年06月08日, 至 2021年06月08日 - DNAに“設計図”以外の新機能を発見 酵素活性を強める効果
東京農工大学大学院が、DNAに「生命の設計図」以外の機能として、酵素活性を強める効果を見つけたと発表したことが紹介される。
yahooニュース／ITmedia／msnニュース／gooニュース／ニコニコニュース／／
自 2021年06月09日, 至 2021年06月09日