Fig. 2

T7 endonuclease I (T7E1) assay and ICE analysis of RTgutGC transfected cells. a Left panel shows a schematic representation of the T7E1 assay. Right panel shows the results of the T7E1 assay visualized on an Agilent Bioanalyzer DNA High Sensitivity chip. 1: cyp1a1 from RNP transfected RTgutGC cells; 2: cyp1a1 from untransfected RTgutGC cells; 3: cyp1a3 from transfected RTgutGC cells; 4: cyp1a3 from untransfected RTgutGC cells; 5: T7E1 assay positive control. Uncut and cleaved fragments, corresponding to the cyp1a1 wild type sequence and indels, respectively, are indicated with dashed lines. Gene editing efficiency is reported above each lane as percentage (%). b Sanger sequencing results of wild type (upper panel) and transfected (lower panel) RTgutGC cells. gRNA and PAM position are indicated by the purple and pink boxes, respectively. Cas9 cutting sites are indicated by vertical red lines. c Output of ICE analysis of the RNP transfected cells. The type of insertions and deletions (indels) detected in the analyzed sample and their respective frequencies are indicated on the left side. On the right side, the cyp1a1 wild-type and edited sequences are aligned. Dashed line indicates the Cas9 cutting site. Created with https://biorender.com