Factors to be considered for improving the HDR (IDT webinar)
Factors to be considered for improving the HDR Improved designing Selection of high activity guides Using the other type of Cas – > changing from Cas9 to Cas12a Blocking mutations to prevent re-cutting after HDR Introducing non-complement base in gRNA to reduce the activity of Cas9 Optimize homology arm length Same length but different gene – > give different HDR efficiency In general sense, larger – > better HDR but it will reach plateau, thus we use the lowest length which gives the best HDR (~200 bp) Improved reagent Small molecule to modulate pw Available by the company (blind solution) Modification of repair template to increase stability Web-based tool to design the gRNA www.idtdna.com/HDRdesigntool Info: www.idtdna.com/HDR www.idtdna.com/CRISPR