PerspectiveRNA Events

Cas9 Targeting and the CRISPR Revolution

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Science  16 May 2014:
Vol. 344, Issue 6185, pp. 707-708
DOI: 10.1126/science.1252964

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  1. Targeting a molecular scalpel.

    The CRISPR-Cas9 nuclease, programmed with sgRNA, specifically targets a short DNA sequence “tag” (the PAM) and unzips DNA complementary to the sgRNA to create a sgRNA–target DNA heteroduplex, triggering R-loop formation. Two nuclease domains (RuvC, HNH) each nick one DNA strand, generating a double-strand break. Structurally, the Cas9 recognition (REC) domain interacts with the sgRNA, while the nuclease (NUC) lobe drives interaction with the PAM and target DNA.

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