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Applications of CRISPR: Tips for designing HDR experiments

Discover how type of donor template, template design, and choice of PAM site can affect efficiency of Cas9-mediated homology-directed repair (HDR). Then read the linked application note for detailed, step-wise guidance to maximize HDR rates in your own genome editing experiments.

CRISPR-Cas9 revolutionizes genomics by enabling efficient site-directed genome editing in a wide variety of biological systems. After double-strand breaks are generated by CRISPR-Cas9, mammalian cells use endogenous cellular machinery to repair the broken sites, exemplified by the canonical non-homologous end joining (NHEJ) pathway and homology-directed repair (HDR) pathway [1]. The error-free HDR faithfully copies genetic information from a related DNA sequence, but this process is characteristically inefficient. As a result, while CRISPR-Cas9 mediated gene knock-out via NHEJ proves to be robust in most organisms, seamless insertion of exogenous genetic material by HDR remains a challenge. Read the full article on IDT's website.