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Specific targeting of plasmids with Argonaute enables genome editing

Esyunina, Daria and Okhtienko, Anastasiia and Olina, Anna and Prostova, Maria and Aravin, Alexei A. and Kulbachinskiy, Andrey (2022) Specific targeting of plasmids with Argonaute enables genome editing. . (Unpublished)

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Prokaryotic Argonautes (pAgos) are programmable nucleases involved in cell defense against invading DNA. Recent studies showed that pAgos can bind small single-stranded guide DNAs (gDNAs) to recognize and cleave complementary DNA in vitro. In vivo pAgos preferentially target plasmids, phages and multicopy genetic elements. Here, we reveal that CbAgo nuclease from Clostridium butyricum can be used for genomic DNA cleavage and engineering in bacteria. CbAgo-dependent targeting of genomic loci with plasmid-derived gDNAs promotes recombination between plasmid and chromosomal DNA. Efficient genome cleavage and recombineering depends on the catalytic activity of CbAgo, its interactions with gDNAs, and the extent of homology between plasmid and chromosomal sequences. Specific targeting of plasmids with Argonautes can be used to integrate plasmid-encoded sequences into the chromosome thus enabling genome editing. One-Sentence SummaryProkaryotic Argonaute nuclease induces DNA interference between plasmid and chromosomal DNA to promote genome recombineering.

Item Type:Report or Paper (Discussion Paper)
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URLURL TypeDescription Paper
Esyunina, Daria0000-0002-3706-4425
Olina, Anna0000-0003-1844-2109
Aravin, Alexei A.0000-0002-6956-8257
Kulbachinskiy, Andrey0000-0002-2292-6424
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This version posted April 14, 2022. We thank Dr. David Leach for insightful discussions, Dr. Aleksei Agapov for help with figure preparation. The authors have declared no competing interest.
Record Number:CaltechAUTHORS:20220419-766504700
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Official Citation:Specific targeting of plasmids with Argonaute enables genome editing. Daria Esyunina, Anastasiia Okhtienko, Anna Olina, Maria Prostova, Alexei A. Aravin, Andrey Kulbachinskiy. bioRxiv 2022.04.14.488398; doi:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:114369
Deposited By: Tony Diaz
Deposited On:19 Apr 2022 15:29
Last Modified:19 Apr 2022 15:29

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