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Abstract
A two-stage protocol for genome engineering in human induced pluripotent stem cells achieves improved editing efficiency and screening throughput.
- The protocol utilizes CRISPR-Cas9 for generating homozygous, multi-kilobase isogenic haplotypes.
- Challenges with low efficiency of homology-directed repair (HDR) and difficulties in generating HDR templates are addressed.
- The optimization leads to a reduction in turnaround time for genome editing.
- Steps include in silico experimental design, large haplotype deletion, and reconstruction.
- Validation of edited cell lines is part of the protocol.
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