Amplicon sequencing of base editing

We performed multiplex gene editing in T cells to create CAR T cells with additional knock-out of HLA class 1 and HLA class 2 by targeting the B2M gene and the CIITA gene. The frequency of different editing outcomes was analysed by amplicon sequencing of the target site in the B2M gene and the target site in the CIITA gene after targeting with either conventional Streptococcus pyogenes Cas9 or the Streptococcus pyogenes nickase Cas9-derived adenine base editor (ABE8.20m).