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CRISPR-based homology-directed repair (HDR) is an invaluable tool to facilitate specific mutations in a genomic region of interest in research studies. While many methods have been reported for improving HDR efficiency, achieving precise changes via HDR remains a challenge, particularly for large knock-ins. These insertions can be generated via HDR using enzymatically generated donor templates.
What you will learn:
How dsDNA donor templates with novel end-modifications improve the frequency of HDR and reduce homology-independent (blunt) insertion events.
How the addition of Alt-R HDR Enhancer V2, a small molecule compound, can further improve HDR rates.
Design considerations when using large dsDNA templates for successful HDR experiments.