Sox2^{em2.1(Snrpn-EGFP)Jae}

CRISPR/cas9 endonuclease-mediated genome editing in is used target the super enhancer (SE) sequences ~100 kb distal of the SRY (sex determining region Y)-box 2 locus (Sox2) on chromosome 3, as well as Snrpn::eGFP::WPRE::BHGpA::loxP-PGK-Puro-loxP sequences (encoding the minimal Snrpn promoter [including conserved elements between human and mouse and containing the endogenous imprinted DMR region], the enhanced green fluorescent protein (EGFP), a WPRE sequence, a BGH polyadenylation sequence and a loxP-flanked PGK-Puro selection cassette). Junction PCR-SNP analyses identified ES cells with Snrpn::eGFP::WPRE::BHGpA sequences inserted into the Sox2 SE on the CAST chromosome. Those ES cells were transfected with a Cre recombinase-expressing plasmid to remove the PGK-Puro cassette. (J:282716)