Genetic manipulations of nonmodel gut microbes.

Autor: Jin WB; Jill Roberts Institute for Research in Inflammatory Bowel Disease, Weill Cornell Medicine Cornell University New York New York USA.; Friedman Center for Nutrition and Inflammation, Weill Cornell Medicine Cornell University New York New York USA., Guo CJ; Jill Roberts Institute for Research in Inflammatory Bowel Disease, Weill Cornell Medicine Cornell University New York New York USA.; Friedman Center for Nutrition and Inflammation, Weill Cornell Medicine Cornell University New York New York USA.; Joan and Sanford I. Weill Department of Medicine, Gastroenterology and Hepatology Division, Weill Cornell Medicine Cornell University New York New York USA.; Department of Microbiology and Immunology, Weill Cornell Medicine Cornell University New York New York USA.; Immunology and Microbial Pathogenesis Program, Weill Cornell Graduate School of Medical Sciences, Weill Cornell Medicine Cornell University New York New York USA.
Jazyk: angličtina
Zdroj: IMeta [Imeta] 2024 Jun 23; Vol. 3 (4), pp. e216. Date of Electronic Publication: 2024 Jun 23 (Print Publication: 2024).
DOI: 10.1002/imt2.216
Abstrakt: Hundreds of microbiota gene expressions are significantly different between healthy and diseased humans. The "bottleneck" preventing a mechanistic dissection of how they affect host biology/disease is that many genes are encoded by nonmodel gut commensals and not genetically manipulatable. Approaches to efficiently identify their gene transfer methodologies and build their gene manipulation tools would enable mechanistic dissections of their impact on host physiology. This paper will introduce a step-by-step protocol to identify gene transfer conditions and build the gene manipulation tools for nonmodel gut microbes, focusing on Gram-negative Bacteroidia and Gram-positive Clostridia organisms. This protocol enables us to identify gene transfer methods and develop gene manipulation tools without prior knowledge of their genome sequences, by targeting bacterial 16s ribosomal RNAs or expanding their compatible replication origins combined with clustered regularly interspaced short palindromic repeats machinery. Such an efficient and generalizable approach will facilitate functional studies that causally connect gut microbiota genes to host diseases.
Competing Interests: The authors declare no conflict of interest.
(© 2024 The Authors. iMeta published by John Wiley & Sons Australia, Ltd on behalf of iMeta Science.)
Databáze: MEDLINE
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