CRISPR-mediated homology-directed repair in hESC or hiPSC
For a single gene, 2 guide RNA constructs and one ssODN or a gene targeting vector will be designed and generated. A total of ~20 clones/sgRNA will be isolated for PCR analysis of knock-in sequences and the PCR products from up to 3 candidates/sgRNA will be confirmed by sequencing. 2-3 lines chosen by the investigator will be expanded and stocks frozen. The entire project should require 18-20 weeks.