Fig. 7.

Validation of CRISPR/Cas9 editing of the TOP2α E19/I19 5′ SS in K/VP.5 cells by Rsa1 analysis. (A) Schematic representation of the E18 through I19 portion of the TOP2α gene. Red arrows denote the forward (For) and reverse (Rev) primers used for the identification of CRISPR editing of the TOP2α E19/I19 5′ SS using RsaI endonuclease. Purple arrow denotes site where Rsa1 creates double-stranded breaks of CRISPR-edited PCR amplicons (B) Ethidium bromide–stained agarose gel of fractionated Rsa1-treated PCR amplicons from K/VP.5 and K/VP.5/edit-1–3 DNA. The black arrow denotes the parental PCR amplicon. Expected sizes of parental and daughter PCR amplicons are indicated in red.