Structural basis for CRISPR RNA-guided DNA recognition by Cascade

@article{Jore2011StructuralBF,
  title={Structural basis for CRISPR RNA-guided DNA recognition by Cascade},
  author={Matthijs M Jore and Magnus Lundgren and Esther van Duijn and Jelle Bernard Bultema and Edze R. Westra and Sakharam Waghmare and Blake Wiedenheft and {\"U}mit Pul and Reinhild Wurm and Rolf Wagner and Marieke R. Beijer and Arjan Barendregt and Kaihong Zhou and Ambrosius P. Snijders and Mark J. Dickman and Jennifer A. Doudna and Egbert J. Boekema and Albert J. R. Heck and John Oost and Stan J. J. Brouns},
  journal={Nature Structural \&Molecular Biology},
  year={2011},
  volume={18},
  pages={529-536}
}
The CRISPR (clustered regularly interspaced short palindromic repeats) immune system in prokaryotes uses small guide RNAs to neutralize invading viruses and plasmids. In Escherichia coli, immunity depends on a ribonucleoprotein complex called Cascade. Here we present the composition and low-resolution structure of Cascade and show how it recognizes double-stranded DNA (dsDNA) targets in a sequence-specific manner. Cascade is a 405-kDa complex comprising five functionally essential CRISPR… 
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TLDR
The E. coli Cascade-crRNA system exhibits significant flexibility suggesting that this complex can be engineered for applications in genome editing and opening the way for incorporation of site-specific labels in crRNA.
CRISPR immunity relies on the consecutive binding and degradation of negatively supercoiled invader DNA by Cascade and Cas3.
Structural basis for promiscuous PAM recognition in type I–E Cascade from E. coli
TLDR
The 2.45 Å crystal structure of Escherichia coli Cascade bound to a foreign double-stranded DNA target is presented, providing the structural basis for understanding the PAM-dependent directional R-loop formation process.
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DNA unwinding is the primary determinant of CRISPR-Cas9 activity
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Detailed kinetic characterization of the enzyme reveals that DNA binding is reversible, R-loop formation is rate-limiting, and the specificity constant for cleavage is determined through an induced-fit mechanism as the product of the equilibrium binding affinity for DNA and the rate of R- loop formation.
Mechanism of CRISPR-RNA guided recognition of DNA targets in Escherichia coli
TLDR
Underlying mechanisms involved in target-induced conformational changes and residues important in DNA binding and protospacer adjacent motif recognition are revealed and highlighted.
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)…
Conformational Dynamics of DNA Binding and Cas3 Recruitment by the CRISPR RNA-Guided Cascade Complex.
TLDR
It is shown how mapping conformational dynamics onto static 3D-structures adds an additional dimension to the functional understanding of this biological machine.
DNA Unwinding Is the Primary Determinant of CRISPR-Cas9 Activity.
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