Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and c-Myc expression for efficient human iPSC induction and differentiation Journal Article
| Authors: | Papapetrou, E. P.; Tomishima, M. J.; Chambers, S. M.; Mica, Y.; Reed, E.; Menon, J.; Tabar, V.; Mo, Q.; Studer, L.; Sadelain, M. |
| Article Title: | Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and c-Myc expression for efficient human iPSC induction and differentiation |
| Abstract: | Human-induced pluripotent stem cells (hiPSCs) are generated from somatic cells by ectopic expression of the 4 reprogramming factors (RFs) Oct-4, Sox2, Klf4, and c-Myc. To better define the stoichiometric requirements and dynamic expression patterns required for successful hiPSC induction, we generated 4 bicistronic lentiviral vectors encoding the 4 RFs co-expressed with discernable fluorescent proteins. Using this system, we define the optimal stoichiometry of RF expression to be highly sensitive to Oct4 dosage, andwedemonstrate the impact that variations in the relative ratios of RF expression exert on the efficiency of hiPSC induction. Monitoring of expression of each individual RF in single cells during the course of reprogramming revealed that vector silencing follows acquisition of pluripotent cell markers. Pronounced lentiviral vector silencing was a characteristic of successfully reprogrammed hiPSC clones, but lack of complete silencing did not hinder hiPSC induction, maintenance, or directed differentiation. The vector system described here presents a powerful tool for mechanistic studies of reprogramming and the optimization of hiPSC generation. |
| Keywords: | controlled study; protein expression; human cell; genetics; nonhuman; animal cell; mouse; cytology; gene expression; animal experiment; animal model; cell differentiation; physiology; gene vector; epigenesis, genetic; myc protein; genes, myc; kruppel like factor; kruppel-like transcription factors; lentivirus vector; transgene; protein induction; teratoma; pluripotent stem cell; pluripotent stem cells; gene silencing; cell clone; genetic epigenesis; octamer transcription factor 4; transcription factor sox2; octamer transcription factor-3; soxb1 transcription factors; oncogene myc; fluorescent proteins; lentiviral vectors; silencing; stoichiometry; kruppel like factor 4; pou5f1 protein, human; sox2 protein, human; transcription factor sox |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 106 |
| Issue: | 31 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 2009-08-04 |
| Start Page: | 12759 |
| End Page: | 12764 |
| Language: | English |
| DOI: | 10.1073/pnas.0904825106 |
| PUBMED: | 19549847 |
| PROVIDER: | scopus |
| PMCID: | PMC2722286 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 14" - "Export Date: 30 November 2010" - "CODEN: PNASA" - "Source: Scopus" |
Altmetric
Citation Impact
BMJ Impact Analytics
MSK Authors
Related MSK Work