ArticleNucleic acids research2022
Programmable cleavage of linear double-stranded DNA by combined action of Argonaute CbAgo from Clostridium butyricum and nuclease deficient RecBC helicase from E. coli.
Article in Nucleic acids research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 54 citations in OpenAlex.
- Advances in Double-Stranded DNA Targeting Technologies.Exploration (Beijing, China) · 2026Review
- A programmable pAgo nuclease with high activity and specificity for efficient DNA and RNA manipulation.Communications biology · 2025Article
- A Mesophilic Argonaute fromBiomolecules · 2025Article
- Random guide-independent DNA cleavage from the Argonaute of Exiguobacterium sp. AB2.BMC microbiology · 2025Article
- Excessive Existence of Positively Charged Amino Acids Caused Off-Target Recognition in the Seed Region ofInternational journal of molecular sciences · 2025Article
- Multiplexed food-borne pathogen detection using an argonaute-mediated digital sensor based on a magnetic-bead-assisted imaging transcoding system.Nature food · 2025Article
- Characterization of argonaute nucleases from mesophilic bacteria Pseudobutyrivibrio ruminis.Bioresources and bioprocessing · 2024Article
- A conditional protein diffusion model generates artificial programmable endonuclease sequences with enhanced activity.Cell discovery · 2024Article
- Article
- MnChemical science · 2024Article
- Programmable site-specific DNA double-strand breaks via PNA-assisted prokaryotic Argonautes.Nucleic acids research · 2023Article
- Applications of Programmable Endonucleases in Sequence- and Ligation-Independent Seamless DNA Assembly.Biomolecules · 2023Review
- DNA rehybridization drives product release from Cas9 ribonucleoprotein to enable multiple-turnover cleavage.Nucleic acids research · 2023Article
- Genome manipulation by guide-directed Argonaute cleavage.Nucleic acids research · 2023Article
- A Programmable, DNA-Exclusively-Guided Argonaute DNase and Its Higher Cleavage Specificity Achieved by 5'-Hydroxylated Guide.Biomolecules · 2022Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Prokaryotic Argonautes (pAgos) use small nucleic acids as specificity guides to cleave single-stranded DNA at complementary sequences. DNA targeting function of pAgos creates attractive opportunities for DNA manipulations that require programmable DNA cleavage. Currently, the use of mesophilic pAgos as programmable endonucleases is hampered by their limited action on double-stranded DNA (dsDNA). We demonstrate here that efficient cleavage of linear dsDNA by mesophilic Argonaute CbAgo from Clostridium butyricum can be activated in vitro via the DNA strand unwinding activity of nuclease deficient mutant of RecBC DNA helicase from Escherichia coli (referred to as RecBexo-C). Properties of CbAgo and characteristics of simultaneous cleavage of DNA strands in concurrence with DNA strand unwinding by RecBexo-C were thoroughly explored using 0.03-25 kb dsDNAs. When combined with RecBexo-C, CbAgo could cleave targets located 11-12.5 kb from the ends of linear dsDNA at 37°C. Our study demonstrates that CbAgo with RecBexo-C can be programmed to generate DNA fragments with custom-designed single-stranded overhangs suitable for ligation with compatible DNA fragments. The combination of CbAgo and RecBexo-C represents the most efficient mesophilic DNA-guided DNA-cleaving programmable endonuclease for in vitro use in diagnostic and synthetic biology methods that require sequence-specific nicking/cleavage of linear dsDNA at any desired location.
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