Rates of homology directed repair (HDR) after a CRISPR cleavage event are often lower than desired. Here, we describe methods and design strategies to improve HDR rates when using ssDNA donors.

Optimized design of ssDNA donors:gRNA and CRISPR enzyme selection

Donor strand selection

Blocking mutations to prevent re-cleavage

 

Alt-R™HDR Design tool ─ a novel bioinformatics tool for HDR template design

HDR donor modifications ─ addition of chemical modifications to stabilize donors

Alt-R HDR Enhancer V2 ─a new small molecule with greater potency for increasing HDR rates and improved cell viability over previously tested compounds.