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Guide-directed DNA cleavage by a prokaryotic Argonaute protein induces chromosome recombination in Escherichia coli

Huang, Shan and Wang, Kaihang and Mayo, Stephen L. (2021) Guide-directed DNA cleavage by a prokaryotic Argonaute protein induces chromosome recombination in Escherichia coli. . (Unpublished) https://resolver.caltech.edu/CaltechAUTHORS:20211122-174116390

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Abstract

Emerging evidence supports the argument that some prokaryotic argonautes (pAgos) serve as a defensive system against invasion of viruses and plasmids through guide DNAs (gDNAs) directed DNA cleavage. This DNA-guided DNA interference motivates research to induce genomic mutations via pAgo mediated cleavage. Here we demonstrate that CbAgo, a pAgo from Clostridium butyricum, is able to induce chromosomal recombination between direct repeat sequences via its gDNA-directed cleavage in Escherichia coli chromosome. We also show that CbAgo targeting can assist Lambda-Red recombineering in RecA-deficient strain. Our study reveals that cleavage by CbAgo in E. coli chromosome can be mutagenic and suggests its broader application in genetic manipulation.


Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/2021.11.17.469050DOIDiscussion Paper
ORCID:
AuthorORCID
Huang, Shan0000-0002-4436-3327
Wang, Kaihang0000-0001-7657-8755
Mayo, Stephen L.0000-0002-9785-5018
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. Version 1 - November 18, 2021; Version 2 - March 25, 2022. We thank Dr. Alexei A. Aravin, Dr. Andrey Kulbachinskiy, Dr. Daria Esyunina, and Dr. David R. F. Leach for helpful discussions. We thank Rochelle A. Diamond and Jamie Tijerina for their help in flow cytometry experiments. All data necessary to support this paper’s conclusions are available in the supplementary materials. Author Contributions: S.H. designed and carried out experiments, with K.W. and S.L.M. providing guidance. S.H., K.W., and S.L.M. wrote the manuscript. The authors declare no competing financial interests.
DOI:10.1101/2021.11.17.469050
Record Number:CaltechAUTHORS:20211122-174116390
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20211122-174116390
Official Citation:Guide-directed DNA cleavage by a prokaryotic Argonaute protein induces chromosome recombination in Escherichia coli. Shan Huang, Kaihang Wang, Stephen L. Mayo. bioRxiv 2021.11.17.469050; doi: https://doi.org/10.1101/2021.11.17.469050
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:111973
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:22 Nov 2021 18:29
Last Modified:25 Mar 2022 22:49

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