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Structural basis for Cas9 off-target activity (Cell 2022 Switzerland Martin Jinek)

Cas9: complement guide CRISPR RNA (crRNA) by cleaving double-stranded DNA (dsDNA) substrates. In addition, by changing the sequence of the guide RNA (gRNA), the DNA-specific programming of the CRISPR-Cas9 system can also be easily realized.
This paper reports the crystal structure of Cas9 binding to a bona fide off-target substrate, revealing that off-target binding is achieved by a series of non-canonical base-pairing interactions in the heteroduplex directing the off-target
These studies provide a structural basis for the off-target activity of Cas9 and help improve the rational design of guide RNAs and off-target prediction algorithms.