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Structural biology

CRISPR preorders for Cas

Nature Reviews Microbiology volume 13, page 459 (2015)Cite this article

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In type II CRISPR−Cas adaptive immunity systems, a CRISPR RNA (crRNA) forms a hybrid structure with a trans-activating crRNA (tracrRNA) that, together with recognition of a protospacer adjacent motif (PAM), guides the Cas9 endonuclease protein to foreign DNA. Jiang et al. solved a 2.9 Å crystal structure of Streptococcus pyogenes Cas9 in complex with a synthetic guide RNA (sgRNA; a crRNA−tracrRNA fusion). sgRNA binding induced several conformational changes in Cas9; notably, the PAM-interacting domain, which is disordered in the apo form of Cas9, adopted a structured conformation that allowed recognition of PAM. Within the sgRNA itself, the 'seed' sequence, which initiates target binding, had an A-form conformation that favours hybridization with DNA. Together, these findings show how the type II CRISPR−Cas complex is preorganized prior to making contact with target DNA.

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References

  1. Jiang, F. et al. A Cas9–guide RNA complex preorganized for target DNA recognition. Science 348, 1477–1481 (2015)

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  1. Naomi Attar
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Attar, N. CRISPR preorders for Cas. Nat Rev Microbiol 13, 459 (2015). https://doi.org/10.1038/nrmicro3547

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