This video explains the concept of CRISPR-mediated cytosine base editors for inducing targeted nucleotide substitution.
Protocol
1. Creation of BRCA1 variants using CRISPR-mediated base editing tools NOTE: If HAP1-BE3 cell lines is not used, BE3-encoding plasmid DNA can be co-transfected with BRCA1-targeting gRNA. Compared to co-transfection of BE3 and gRNA plasmids, transfection of gRNA plasmid to HAP-BE3 cells induce efficient base editing up to 3-fold at target locus in our hands. Seed 5 x 105 HAP1-BE3 cells (or HAP1 cells …
CRISPR-Mediated Base Editing Tools: A Genome Editing Technique to Induce Targeted Base Substitution. J. Vis. Exp. (Pending Publication), e20970, doi: (2023).