Tg(tetO-Pou5f1,-Sox2,-Klf4,-Myc)1Srn

The Tet-O-FUW-OSKM lentiviral vector was designed with a tetracycline responsive element (TRE, tetOP, or tetO) and a minimal CMV promoter driving expression of four mouse genes: Oct4 (Pou5f1; POU domain, class 5, transcription factor 1), Klf4 (Kruppel-like factor 4 (gut)), Sox2 (SRY-box containing gene 2), and c-Myc (Myc; myelocytomatosis oncogene). Expression of Pou5f1, Klf4, Sox2, and Myc were separated by three different viral 2A oligopeptides that mediate ribosomal skipping (P2A [from porcine teschovirus-1], T2A [from insect Thosea asigna virus], and E2A [equine rhinitis A virus], respectively). C57BL/6-derived mouse embryonic fibroblasts (MEFs), carrying a doxycycline-inducible transcriptional activator (rtTA) within the Rosa26 locus (see Stock No. 006965) were transduced with the lentivirus Tet-O-FUW-OSKM. Treatment of MEFs with doxycycline reprogrammed the MEFs into induced pluripotent stem cells (iPSCs). The lentiviral transgenes were transmitted at Mendelian proportions, indicative of single integration site. Inducible four factors lines i4F-A (also known as line 1) and i4F-B were generated. In this line, the transgene integrated within the fourth intron of the Neto2 gene, which resides on Chr 8. (J:201192)