Fig. 1
TALEN and gRNA design, and comparison of these gene editors on EGFP gene disruption. a Schematic diagram of target sites for TALENs and gRNAs proximal to the two substitution sites (199T > C) and (437A > T) for conversion of EGFP to EBFP. Coloured arrow lines indicate the sequence used for the guide segment of gRNAs. The NGG nucleotide protospacer adjacent motif (PAM) sequences in red. Colour boxed sequences denote the DNA binding regions of the TALEN proteins. b Surveyor nuclease assay of genomic DNA isolated from HEK293FTEGFP cells with one copy of integrated EGFP gene expressing Cas9 and gRNAs, or TALENs. NC: genomic DNA from untransfected HEK293FTEGFP cells was used a negative control. c The representatives of examination of EGFP negative cells in HEK293FTEGFP cells transfected with Cas9 and gRNAs, or TALENs under a phase-contrast and fluorescence microscope, and quantification of EGFP negative cells using by fluorescence-activated cell sorter (FACS). Arrow heads indicate EGFP negative cells. Scale bar of 50 μm. For FACS analysis, in each case, a total of 10,000 events were counted