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Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage.

  • Author(s): Jiang, Fuguo
  • Taylor, David W
  • Chen, Janice S
  • Kornfeld, Jack E
  • Zhou, Kaihong
  • Thompson, Aubri J
  • Nogales, Eva
  • Doudna, Jennifer A
  • et al.

Published Web Location

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5111852/
No data is associated with this publication.
Abstract

Bacterial adaptive immunity and genome engineering involving the CRISPR (clustered regularly interspaced short palindromic repeats)-associated (Cas) protein Cas9 begin with RNA-guided DNA unwinding to form an RNA-DNA hybrid and a displaced DNA strand inside the protein. The role of this R-loop structure in positioning each DNA strand for cleavage by the two Cas9 nuclease domains is unknown. We determine molecular structures of the catalytically active Streptococcus pyogenes Cas9 R-loop that show the displaced DNA strand located near the RuvC nuclease domain active site. These protein-DNA interactions, in turn, position the HNH nuclease domain adjacent to the target DNA strand cleavage site in a conformation essential for concerted DNA cutting. Cas9 bends the DNA helix by 30°, providing the structural distortion needed for R-loop formation.

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