xCas9: Engineering a CRISPR Variant with PAM Flexibility
Type
Blog Post
Published
March 28, 2018, 6:52 p.m.
...recognizing a broad range of PAMs like NG, GAA, and GAT, but also displaying increased editing specificity...showed the broadest PAM range, recognizing NG, NNG, GAT, and CAA PAMs, while xCas9 3.6 displayed the second...at an NGG PAM. When they tested a variety of NG, GAT, GAA, and CAA PAMs, they saw that xCas9 3.7 cleaves...efficiency), NGC (2.1-fold), NGA (1.6-fold) and GAA and GAT (5.2-fold). The low increase in NGA editing efficiency...activating transcription. Sites containing NG, GAA, or GAT PAMs averaged 56-91% transcriptional activation compared...28%). Cytidine base editing at NGT, NGC, GAA, and GAT PAMs is improved, with a smaller increase in NGA ...containing ABE7.10, as well as improvements at NG and GAT PAMs.
xCas9 3.7 Activity in Mammalian Cells
Construct...