Fig. 2

Construction and verification of an endogenous GFP Nanog marker. a) Map of the CRISPR/Cas9 target site on exon 4 of the Nanog gene, the gRNA sequence. b) The PCR GFP products of the A2780 + 20 and SKOV3 + 5 GFP (+) clonal cells (agarose gel electrophoresis). The negative control is the non-targeted A2780 cells without GFP. c) Enzyme digestion of PCR products from A2780 + 20 and SKOV3 + 5 monoclonal cells. d) The Sanger sequencing results: the bases labeled in blue represent the end of the Nanog exon 4 and the first of the 2A joint sequences in A2780 + 20 and SKOV3 + 5 monoclonal cells