CRISPR 101: Cas9 Nickase Design and Homology Directed Repair
Type
Blog Post
...middle of the targeted region, as shown below.
To identify optimal nickase designs, IDT scientists Mollie...insert an EcoRI site using a single-stranded oligonucleotide (ssODN) donor. Using a D10A nickase to insert...nickase outperformed both WT Cas9 sites tested by a wide margin, exhibiting >20% repair efficiency. Clearly... is especially useful if there are no available guides close to the desired mutation site.
Quick tips...small insertions or tags (ssODN; IDT Ultramer® Oligonucleotides)
Use 100 nt homology arms for large insertions...
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