Tn(pb-CAG-cas9*,-EGFP)1Yangh

The targeting vector contains a CAG promoter, floxed STOP cassette, dCas9 sequence, HA tag, 10 copies of the GCN4 (General Control Protein) epitope peptide sequence, P2A, scFv sequence encoding cognate single-chain antibody that recognizes GCN4, P65-HSF1 activator helper complex, T2A, EGFP, and a woodchuck hepatitis post transcriptional regulatory element (WPRE). This construct was flanked by piggyBac transposase inverted terminal repeats and was co-injected with a plasmid encoding piggyBac transposase expressed under the control of a beta-actin promoter. Genetic analysis at The Jackson Laboratory indicates that one copy of the transgene inserted on chromosome 14 within Dpysl2 intron 3, rather than than the Gm33925 locus as originally reported. It is not known if the transgene insertion has any effect on Dpysl2 expression. (J:259469)