Lipid-mediated transfection of CRISPR-Cas9 and single-strand DNA oligos for homology-directed repair
The Dharmacon Edit-R CRISPR-Cas9 Genome Engineering platform has previously been demonstrated to easily create insertions and deletions (indels) to disrupt genes using Cas9, synthetic tracrRNA and a synthetic crRNA. In this poster, we demonstrate that this same system can create precise insertions using the homology-directed repair (HDR) machinery when we supply the cells with a single-strand DNA donor.
- Demonstration of precise insertions with efficiencies up to 25%
- Insertion of 10-12 nucleotide sequences with optimized DNA donor oligo homology arm length
- Simple and straight forward experimental workflow for HDR lipid mediated transfections
The Edit-R CRISPR-Cas9 gene engineering platform is a new and exciting molecular tool to interrogate gene function. We are pleased to provide this resource demonstrating how it can be used as part of an HDR lipid transfection workflow to create precise knockins.
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