How do I design my HDR template?
How do I design my HDR template?
The sequence of the HDR template should match the intended sequence of the targeted genomic locus, including the desired edit flanked by 5' and 3' ends that share homology with the wild-type genomic locus. (These 5' and 3' sequences are referred to as ""homology arms."")
Sequences corresponding to desired edits (insertions, substitutions, etc.) should be positioned in the middle of the HDR template. The exact sequence of the HDR template is independent of the position of the single guide RNA (sgRNA) or the cut site of the Cas9 nuclease, but efficient editing requires a template design in which the sequence(s) to be inserted are in relatively close proximity to the cut site of the Cas9-sgRNA ribonucleoprotein (RNP).