DNA-guided DNA cleavage at moderate temperatures by Clostridium butyricum Argonaute
Jorrit W. Hegge (Wageningen University & Research)
Daan C. Swarts (Universitat Zurich, Wageningen University & Research)
S.D. Chandradoss (Kavli institute of nanoscience Delft, TU Delft - BN/Chirlmin Joo Lab)
Tao Ju Cui (Kavli institute of nanoscience Delft, TU Delft - BN/Chirlmin Joo Lab)
Jeroen Kneppers (Wageningen University & Research)
Martin Jinek (Universitat Zurich)
C. Joo (TU Delft - BN/Chirlmin Joo Lab, Kavli institute of nanoscience Delft)
John Van Der Oost (Wageningen University & Research)
More Info
expand_more
Other than for strictly personal use, it is not permitted to download, forward or distribute the text or part of it, without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license such as Creative Commons.
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.
help_outline