Here, we report a rapid CRISPR-Cas9-mediated gene knock-in strategy that uses Cas9 ribonucleoprotein and 5'-modified double-stranded DNA donors with 50-base-pair homology arms and achieved unprecedented 65/40% knock-in rates for 0.7/2.5 kilobase inserts, respectively, in human embryonic kidney 293T cells. The identified 5'-end modification led to up to a fivefold increase in gene knock-in rates at various genomic loci in human cancer and stem cells.
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|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7085973 | PMC |
| http://dx.doi.org/10.1038/s41589-019-0432-1 | DOI Listing |
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