A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity

@article{Jinek2012APD,
  title={A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity},
  author={Martin Jinek and Krzysztof Chylinski and Ines Fonfara and Michael H. Hauer and Jennifer A. Doudna and E. Charpentier},
  journal={Science},
  year={2012},
  volume={337},
  pages={816 - 821}
}
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. [] Key Result At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand.
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TLDR
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TLDR
Atomic resolution structures of a full-length Cas3 are shown, revealing how Cas3 coordinates binding, ATP-dependent translocation, and nuclease digestion of invader DNA and revealing important mechanistic details on the neutralization of genetic invaders by type I CRISPR-Cas systems.
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TLDR
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TLDR
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CRISPR-Cas9 Structures and Mechanisms.
TLDR
This review aims to provide an in-depth mechanistic and structural understanding of Cas9-mediated RNA-guided DNA targeting and cleavage and provides a framework for rational engineering aimed at altering catalytic function, guide RNA specificity, and PAM requirements and reducing off-target activity for the development of Cas 9-based therapies against genetic diseases.
CasA mediates Cas3-catalyzed target degradation during CRISPR RNA-guided interference
TLDR
Together, these data show that the CasA subunit of Cascade functions as an essential partner of Cas3 by recognizing DNA target sites and positioning Cas3 adjacent to the PAM to ensure cleavage, and biochemically that base pairing of the Pam region is unnecessary for target binding but critical for Cas3-mediated degradation.
CRISPR‐Cas systems and RNA‐guided interference
TLDR
A number of studies have shown that CRISPR‐mediated immunity can readily increase the breadth and depth of virus resistance in bacteria and archaea and applications of crRNA‐guided interference are discussed.
Mechanism of foreign DNA recognition by a CRISPR RNA-guided surveillance complex from Pseudomonas aeruginosa
The Type I-F CRISPR-mediated (clustered regularly interspaced short palindromic repeats) adaptive immune system in Pseudomonas aeruginosa consists of two CRISPR loci and six CRISPR-associated (cas)
A Cas9–guide RNA complex preorganized for target DNA recognition
TLDR
The guide RNA in the CRISPR-Cas immune/editing system is poised to initiate recognition of target DNA and is construe this as convergent evolution of a “seed” mechanism reminiscent of that used by Argonaute proteins during RNA interference in eukaryotes.
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