DNA-guided DNA cleavage at moderate temperatures by Clostridium butyricum Argonaute

Jorrit W. Hegge, Daan C. Swarts, Stanley D. Chandradoss, Tao Ju Cui, Jeroen Kneppers, Martin Jinek, Chirlmin Joo, John van der Oost*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

Prokaryotic Argonaute proteins (pAgos) constitute a diverse group of endonucleases of which some mediate host defense by utilizing small interfering DNA guides (siDNA) to cleave complementary invading DNA. This activity can be repurposed for programmable DNA cleavage. However, currently characterized DNA-cleaving pAgos require elevated temperatures (≥65°C) for their activity, making them less suitable for applications that require moderate temperatures, such as genome editing. Here, we report the functional and structural characterization of the siDNA-guided DNA-targeting pAgo from the mesophilic bacterium Clostridium butyricum (CbAgo). CbAgo displays a preference for siDNAs that have a deoxyadenosine at the 5'-end and thymidines at nucleotides 2-4. Furthermore, CbAgo mediates DNA-guided DNA cleavage of AT-rich double stranded DNA at moderate temperatures (37°C). This study demonstrates that certain pAgos are capable of programmable DNA cleavage at moderate temperatures and thereby expands the scope of the potential pAgo-based applications.

Original languageEnglish
Pages (from-to)5809-5821
Number of pages13
JournalNucleic acids research
Volume47
Issue number11
DOIs
Publication statusPublished - 20 Jun 2019

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Clostridium butyricum
DNA Cleavage
Argonaute Proteins
Temperature
DNA
Endonucleases
Thymidine
Nucleotides
Complementary DNA
Bacteria

Cite this

Hegge, Jorrit W. ; Swarts, Daan C. ; Chandradoss, Stanley D. ; Cui, Tao Ju ; Kneppers, Jeroen ; Jinek, Martin ; Joo, Chirlmin ; van der Oost, John. / DNA-guided DNA cleavage at moderate temperatures by Clostridium butyricum Argonaute. In: Nucleic acids research. 2019 ; Vol. 47, No. 11. pp. 5809-5821.
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abstract = "Prokaryotic Argonaute proteins (pAgos) constitute a diverse group of endonucleases of which some mediate host defense by utilizing small interfering DNA guides (siDNA) to cleave complementary invading DNA. This activity can be repurposed for programmable DNA cleavage. However, currently characterized DNA-cleaving pAgos require elevated temperatures (≥65°C) for their activity, making them less suitable for applications that require moderate temperatures, such as genome editing. Here, we report the functional and structural characterization of the siDNA-guided DNA-targeting pAgo from the mesophilic bacterium Clostridium butyricum (CbAgo). CbAgo displays a preference for siDNAs that have a deoxyadenosine at the 5'-end and thymidines at nucleotides 2-4. Furthermore, CbAgo mediates DNA-guided DNA cleavage of AT-rich double stranded DNA at moderate temperatures (37°C). This study demonstrates that certain pAgos are capable of programmable DNA cleavage at moderate temperatures and thereby expands the scope of the potential pAgo-based applications.",
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DNA-guided DNA cleavage at moderate temperatures by Clostridium butyricum Argonaute. / Hegge, Jorrit W.; Swarts, Daan C.; Chandradoss, Stanley D.; Cui, Tao Ju; Kneppers, Jeroen; Jinek, Martin; Joo, Chirlmin; van der Oost, John.

In: Nucleic acids research, Vol. 47, No. 11, 20.06.2019, p. 5809-5821.

Research output: Contribution to journalArticleAcademicpeer-review

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