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| Item Type: | Article |
|---|---|
| Title: | Novel codon-optimized mini-intronic plasmid for efficient, inexpensive, and xeno-free induction of pluripotency |
| Creators Name: | Diecke, S., Lu, J., Lee, J., Termglinchan, V., Kooreman, N.G., Burridge, P.W., Ebert, A.D., Churko, J.M., Sharma, A., Kay, M.A. and Wu, J.C. |
| Abstract: | The development of human induced pluripotent stem cell (iPSC) technology has revolutionized the regenerative medicine field. This technology provides a powerful tool for disease modeling and drug screening approaches. To circumvent the risk of random integration into the host genome caused by retroviruses, non-integrating reprogramming methods have been developed. However, these techniques are relatively inefficient or expensive. The mini-intronic plasmid (MIP) is an alternative, robust transgene expression vector for reprogramming. Here we developed a single plasmid reprogramming system which carries codon-optimized (Co) sequences of the canonical reprogramming factors (Oct4, Klf4, Sox2, and c-Myc) and short hairpin RNA against p53 ("4-in-1 CoMiP"). We have derived human and mouse iPSC lines from fibroblasts by performing a single transfection. Either independently or together with an additional vector encoding for LIN28, NANOG, and GFP, we were also able to reprogram blood-derived peripheral blood mononuclear cells (PBMCs) into iPSCs. Taken together, the CoMiP system offers a new highly efficient, integration-free, easy to use, and inexpensive methodology for reprogramming. Furthermore, the CoMIP construct is color-labeled, free of any antibiotic selection cassettes, and independent of the requirement for expression of the Epstein-Barr Virus nuclear antigen (EBNA), making it particularly beneficial for future applications in regenerative medicine. |
| Keywords: | Cellular Reprogramming, Codon, Cultured Cells, Epstein-Barr Virus Nuclear Antigens, Fibroblasts, Fluorescence Microscopy, Homeodomain Proteins, Innate Immunity, Introns, Karyotyping, Mononuclear Leukocytes, Plasmids, Pluripotent Stem Cells, Proto-Oncogene Proteins c-myc, RNA-Binding Proteins, Small Interfering RNA, Transcription Factors, Tumor Suppressor Protein p53, Animals, Mice |
| Source: | Scientific Reports |
| ISSN: | 2045-2322 |
| Publisher: | Nature Publishing Group |
| Volume: | 5 |
| Page Range: | 8081 |
| Date: | 28 January 2015 |
| Official Publication: | https://doi.org/10.1038/srep08081 |
| PubMed: | View item in PubMed |
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