xCas9: Engineering a CRISPR Variant with PAM Flexibility
Type
Blog Post
...targeting anywhere in an exon. But things get more complicated if you’d like to use homology directed repair...alternative PAM sequences are longer and often more complex than NGG, resulting in lower overall genome frequency...Characterizing xCas9 in mammalian cells
Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells...PAMs averaged 56-91% transcriptional activation compared to the NGG PAM, confirming the flexibility of ...with improved editing at an NGG PAM (69 vs 48%) compared to SpCas9-containing ABE7.10, as well as improvements...NGS) are shown as percentages, with fold changes compared to SpCas9 in parentheses.
The development of ...that selecting for one beneficial trait, PAM compatibility, also produced an enzyme with improved on-target...