Chapter 4 —— 67 —— Figure 7. Investigating off-target AAV donor insertions resulting from using regular versus high-specificity CRISPR-Cas9 complexes. (A) Experimental setup for experiments in hMSCs. AAV HR donor sequences before and after chromosomal insertion at AAVS1 and off-target CPNE5 loci through HDR-dependent and HDR-independent processes, respectively, are shown. Likewise for the respective amplicons specific for ontarget and off-target insertions. In addition to AAV-HRS1, hMSCs were exposed to high-
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