FIGURE 3.

Frequency and pattern of gene modifications following ZFN administration in mouse neurons. (A) On-target and off-target indel frequencies for the CatD gene after 2 and 3 weeks of ZFN expression. Amplicons for the target sequence of CatD were obtained from the DNA isolated from the neurons transduced with ZFN against CatD. Neurons transduced with the ZFN1 and ZFN2 individually and those with non-CatD ZFN served as negative control. Off-target amplicons were obtained from the DNA collected from the neurons expressing CatD ZFN. (B) Distribution of indel length (Number of deletions and insertions) in CatD gene after 2 and 3 weeks of ZFN expression. For (A,B), error bars reflect SEM from three biological replicates performed on independent experiments. (C) Representative examples of genomic DNA sequences at CatD locus that are modified following ZFN transduction. The unmodified genomic site is the first sequence, followed by the most abundant sequences containing deletions and insertions. The stars represent gaps, a dot for a match on the same strand.