CRISPR-Mediated Base Editing Tools: A Genome Editing Technique to Induce Targeted Base Substitution

Seed 5 x 10⁵ HAP1-BE3 cells per well in 24-well plates one day prior to transfection, and culture them to reach 70% to 80% confluence for transfection. Transfect BRCA1-targeting guide RNAs using the purchase transfection reagents, according to the manufacturer's protocol. Use 1 microgram of BRCA1 targeting guide RNAs to induce CG to TA conversion at BRCA1 target sites. Then, incubate the cells at 37 degrees Celsius and subculture every 3 to 4 days. Harvest the cell pellets 3, 10, and 24 days after transfection to analyze base editing efficiencies. Extract genomic DNA using the genomic DNA purification kit.