| 研究課題/領域番号 |
22K20894
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| 研究種目 |
研究活動スタート支援
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| 配分区分 | 基金 |
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| 審査区分 |
0902:内科学一般およびその関連分野
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| 研究機関 | 自治医科大学 |
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研究代表者 |
Nguyen・Minh・Thuy 自治医科大学, 医学部, ポスト・ドクター (20964191)
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| 研究期間 (年度) |
2022-08-31 – 2024-03-31
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| 研究課題ステータス |
完了 (2023年度)
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| 配分額 *注記 |
2,860千円 (直接経費: 2,200千円、間接経費: 660千円)
2023年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
2022年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
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| キーワード | CRISPR-Cas13a / Bacteriophage / gut microbe / Bacteroides fragilis / Identification |
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| 研究開始時の研究の概要 |
Enterotoxigenic Bacteroides fragilis (ETBF) is the cause of inflammatory diarrhea of the colon and intestinal cancer. 5-20% of the world population has bft gene-positive asymptomatic ETBF detected in the gut microbiota. In this study, we developed an identification system that selectively kills and identifies ETBF by introducing CRISPR-Cas13a, which targets the bft toxin gene, into the phage capsid. The results of this application establish a fast and easy genotyping method that does not require nucleic acid amplification.
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| 研究実績の概要 |
Enterotoxigenic Bacteroides fragilis (ETBF) is the cause of inflammatory diarrhea of the colon and intestinal cancer. 5-20% of the world population has bft gene-positive asymptomatic ETBF detected in the gut microbiota. In this study, We induced prophages (lysogenic phages) from the bacterial genome using mitomycin C against B. fragilis 310 strains, and isolated the prophages obtains total 5 sample. The infection rates of the isolated prophages 5 strains were determined using clinical isolates of B. fragilis 73 strains (1.36 %-5.47%). The phage morphology was also observed using a transmission electron microscope (TEM) and identified by whole genome analysis. Next, we searched for and optimized CRISPR-cas sequences that recognize the target gene: Using the sequence of the bft gene, we designed a guide RNA that recognizes the bft gene in the 39 bft-carrying strain? and examined its bactericidal activity. We are currently constructing AB capsids using the best candidates. Furthermore, we created a B. fragilis-infected model mouse and constructed an in vivo evaluation system for the bactericidal activity of the target bacteria.
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