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The CRISPR-associated genome editor nuclease Cas9's target DNA specificity is determined by complementarity to a 20-nucleotide segment in its guide RNA. Here we report crystallographic profiles of Cas9 bound to bona fide off-target substrates, revealing that off-target binding is facilitated by a variety of non-canonical base pairing interactions within the guide-off-target heteroduplex. Base skipping or multiple non-canonical base pairs are preferred over RNA bulge formation for off-target sites with single-nucleotide deletions relative to the guide RNA.
Source link: https://doi.org/10.1101/2021.11.18.469088
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