Methods Of Genome Editing Including Controlled Opening Of Chromatin
Inventors: Krishanu Saha, Kaivalya Molugu
UW-Madison researchers developed an improved method of gene editing human iPSCs through CRISPR-Cas. They added the histone deacetylase small molecule inhibitor Trichostatin A (TSA) before addition of the CRISPR-Cas machinery to prime the chromatin before the editing and observed that TSA increased the gene-editing efficiency in iPSCs by ∼3.5 fold. Concentration of TSA affected cell viability (> 25 ng/ml of TSA led to a significant decrease in cell viability), so the inventors used TSA concentrations of 0, 3.13, 6.25, and 12.5 ng/mL in their studies. The inventors used the Saha RNP complex for the gene editing work.
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