Efficient Site-Specific Integration Of New Genetic Information Into Human Cells

Patent Status

Brief Description

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.

Suggested uses

• Genome editing (e.g., human, plants, and animal cells)
• Genome engineering in both transformed and primary human cells

Advantages

• Enhanced HDR efficiency in human cells
• Highly efficient, site-specific editing
• Simple and robust editing
• Cell mortality minimized

Publication

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