Establishment of a novel identification system for Enterotoxigenic Bacteroides fragilis using CRISPR-Cas13 and bacteriophage technology

利用CRISPR-Cas13和噬菌体技术建立产肠毒素脆弱拟杆菌的新型鉴定系统

• 批准号: 22K20894
• 负责人: Nguyen・Minh・Thuy
• 金额: $ 1.83万
• 依托单位: Jichi Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-08-31 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22K20894/
• 关键词: CRISPR-Cas13a; Bacteriophage; gut microbe; Bacteroides fragilis; Identification

Enterotoxigenic Bacteroides fragilis (ETBF) is the cause of inflammatory diarrhea of the colon and intestinal cancer. 5-20% of the world's population carries this strain asymptomatically. Therefore, rapid and easy detection methods are needed to properly prevent and treat the disease. Our laboratory has independently developed an antibacterial capsid using the CRISPR-Cas13a system. This application aims to establish a simple identification system using the bft gene of Bacteroides fragilis. This study culminates in the development of new therapeutic strategies for controlling gut microbiota in the gut microbiota community.In the first year, I have achieved some results: I collected 215 Bacteroides fragilis (BF) strains, including 12 bft1-positive, 8 bft-2-positive, and 1 bft-3-positive strain. I performed whole-genome sequencing on 50 strains, completing a comprehensive survey of their genomes and found out 11 strains revealed prophage sequences in their genome. Based on the results of MIC, six strains that susceptible to antibiotics and not carry prophage in their genome, were suitable to carry Capsid.

产肠毒素脆弱类杆菌（ETBF）是结肠炎性腹泻和肠癌的原因。世界上5 - 20%的人口无症状地携带这种菌株。因此，需要快速简便的检测方法来正确预防和治疗该疾病。我们实验室使用CRISPR-Cas13a系统独立开发了一种抗菌衣壳。本申请旨在利用脆弱类杆菌bft基因建立一个简单的鉴定系统。这项研究的高潮是开发新的治疗策略，以控制肠道菌群中的肠道菌群。在第一年，我取得了一些成果：我收集了215个脆弱类杆菌（BF）菌株，其中12个bft 1阳性，8个bft-2阳性，1个bft-3阳性菌株。我对50个菌株进行了全基因组测序，完成了对其基因组的全面调查，发现11个菌株在其基因组中显示了原噬菌体序列。根据MIC结果，6株对抗生素敏感且基因组中不携带原噬菌体的菌株适合携带衣壳蛋白。

项目成果

Development of Cas13a-phagecapsid to eliminate enterotoxigenic Bacteroides fragilis.

开发 Cas13a-噬菌体衣壳以消除产肠毒素脆弱拟杆菌。

• 发表时间: 2023
• 作者: 10.Thuy Nguyen，Arbaah Mahmoud，相羽 由詞，渡邊 真弥，宮永 一彦，Xin-Ee Tan，笹原 鉄平，崔 龍洙