|United States Of America||Issued Patent||10,570,418||02/25/2020||2014-199|
The CRISPR/Cas9 system is a robust genome editing technology that works based on the RNA-programmed DNA cleaving activity of the Cas9 enzyme. Cas9 generates blunt double-strand DNA (dsDNA) breaks (DSBs) at sites defined by a guide sequence contained within an associated CRISPR RNA (crRNA) transcript and has transformed the ability to engineer eukaryotic organisms by initiating DNA repair pathways that lead to targeted genetic re-programming. There is a need, however, for methods that increase the efficiency of target DNA modification by Cas9 targeting complexes and/or increase the effectiveness of such methods.
UC Berkeley researchers have found a simple and robust approach for introducing new genetic information site-specifically, in a eukaryotic cell, and with high efficiency using timed delivery of Cas9-guide RNA ribonucleoprotein (RNP) complexes. The approach combines well-established cell cycle synchronization techniques with direct nucleofection of the pre-assembled Cas9 ribonucleoprotein (RNP) complexes to achieve controlled nuclease action at the phase of the cell cycle best for HDR.
Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery @import url(https://ottwebapps.ucop.edu/NCD/CuteSoft_Client/CuteEditor/Load.ashx?type=style&file=SyntaxHighlighter.css);
Genetic engineering, homology-directed repair, HDR, genome editing, Cas9, CRISPR