Figure 4.

megaTAL addressing selectively increases on-target cleavage. (a) I-AniI human genomic near-native sequences and their location in the genome. Synthetic TAL effectors were built against the +9T and +5A+8T sequences underlined in green and the I-AniI near-natives sequences are underlined in blue. (b) Level of gene targeting in TLR cells with endogenous human I-AniI near-native targets using ‘addressed’ +9T and +5A+8T megaTALs and ‘unaddressed’ I-AniI mns. mutNHEJ was not measured via TLR due to the presence of a stop codon in some target sites that prevented flow cytometry readout of these events. (c) Mutation rates at endogenous targets using ‘addressed’ I-AniI megaTALs and ‘unadressed’ mns. The level of mutNHEJ at the endogenous I-AniI near-native targets in 293T cells after 72 h of treatment with mn and megaTAL variants was determined based on MiSeq sequencing results.