...spacing
D10A: nick sites separated by 37-68 bp
H840A: nick sites separated by 51-68 bp
Use D10A for HDR...use SpCas9 nickases as examples for this post. The D10A mutation inactivates the RuvC domain, so this nickase...preferences using HEK293 cells. They started by comparing D10A and H840A total editing efficiency in PAM-in and...higher in the PAM-out configuration. In addition, D10A editing efficiency is consistently higher than that...H840A, especially with smaller nick distances. Using D10A, they subsequently showed that editing efficiency...gRNAs are too close (7-23 nt nick distance).
Both D10A and H840A are potent editors, but their indel profiles...profiles vary. In the PAM-out configuration, D10A tends to generate small deletions, while H840A is biased...