Figure 2: Optimization of targeted deamination frequency of AID fusions in E.coli.
From: Engineering and optimising deaminase fusions for genome editing
(a) Schematic representation of ZF-AIDs variants tested for targeted deaminase activity (upper) and the reporter (lower) with the ZF-recognition sequence in blue. (b) GFP rescue by expression of the four ZF-AIDs variants and ZF or AID domains alone. All error bars indicate s.d. (c) Schematic representation of TALE-AIDs and the reporters tested for targeted deaminase activity. Five TALE-AIDs (upper) with different TALE C-terminus truncations (C1–C5) were constructed, with the remaining C-terminus lengths shown in parentheses. Full TALE-AID protein sequences can be found in Supplementary Note 2. Five reporters were constructed (lower) with different spacer lengths (2, 5, 8 and 11 bp) between the broken start codon and TALE DNA binding motif. The TALE binding site on the GFP reporter is shown in blue; the TALE N-terminus segment specifies the 5′ thymine base of the binding site. (d) All five TALE-AIDs were tested for targeted deaminase activity on all five reporters (c). Green and grey encode high and low GFP rescue, respectively.
