Authors:
Cesar A. Sommer, Matthias Stadtfeld, George J. Murphy, Konrad Hochedlinger, Darrell N. Kotton, & Gustavo Mostoslavsky
Summary:
Induced Pluripotent Stem (iPS) cells can be generated using retroviral vectors expressing Oct4, Klf4, Sox2 and cMyc. Most prior studies have required multiple retroviral vectors for reprogramming, resulting in high numbers of genomic integrations in iPS cells and limiting their use for therapeutic applications. Here we describe the use of a single lentiviral vector expressing a ‘stem cell cassette' comprised of the four transcription factors and a combination of 2A peptide and IRES technology, generating iPS cells from post-natal fibroblasts. iPS cells generated in this manner display ES cell-like morphology, express stem cell markers and exhibit in vivo pluripotency as evidenced by their ability to differentiate in teratoma assays and their robust contribution to mouse chimeras. Combining all factors into a single transcript achieves the most efficient reprogramming system to date, and allows derivation of iPS cells with a single viral integration. The use of a single lentiviral vector for reprogramming represents a powerful laboratory tool and a significant step toward the application of iPS technology for clinical purposes.
Source:
Stem Cells; (12/18/08)