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. 2014 Feb 11;9(2):e88146. doi: 10.1371/journal.pone.0088146

Figure 1. TALEN design and evaluation of cutting efficiency in rat glioma C6 cells.

Figure 1

(a) Schematic of the rat Nc3r1 (GR) gene. Zoom on the area of the mutation pA476T in exon 3. The first nucleotide of the 476 codon is highlighted in blue. TALEN binding sites of TAL 3 are highlighted in green. Detailed sequences of TAL 6 binding sites can be found in Table S1, File S1. (b) T-endo1 assay results. Pooled DNA from C6 cells transfected with either Right, Left or Right and Left TALEN monomers (marked with R, L or RL, respectively) was amplified and treated with T7 endo 1 enzyme. Cut bands of 288 and 177 bp indicate TALEN activity. Mcells are mock transfected cells, GFP: GFP transfected cells were used as a positive transfection control. Intensity of the cut bands are indicated for TAL 3 and TAL 6 pairs. (c) TAL 3 transfected cells screening. PCR amplicons of the region around the pA76T mutation were subcloned into TOPO vector. Clones were isolated and analyzed individualy. Four point mutations and insertion.s are marked in red.