xCas9: Engineering a CRISPR Variant with PAM Flexibility
Type
Blog Post
Published
March 28, 2018, 6:52 p.m.
...work, there are other options. Commonly used Cpf1 variants, with a 5’ TTTV PAM, improve targeting in AT-...respectively. Researchers also engineered SpCas9 variants with alternative PAMs like NGAG and NGCG and SaCas9... in the pool. According to this logic, ω-dCas9 variants recognizing multiple PAMs should have a fitness...promoting survival of only the broadest-targeting variants. For the final round of evolution, they used a... a no-G PAM library to enrich for Cas9 variants with activity on non-NGG PAMs, generating xCas9 3.0-3.13...confirming the flexibility of xCas9 3.7.
Base editor variants with xCas9 are similarly flexible. Like the xCas9...References
Hu, Johnny H., et al. "Evolved Cas9 variants with broad PAM compatibility and high DNA specificity...