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Induced Pluripotent Stem Cell Generation Using a Single Lentiviral Stem Cell Cassette
Open accessAbstract
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” composed of the four transcription factors and a combination of 2A peptide and internal ribosome entry site technology, generating iPS cells from postnatal fibroblasts. iPS cells generated in this manner display embryonic stem 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.
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“Polycistronic technology reduces the requirement for multiple viral vectors to deliver four different transcription factors, cutting this down to a single-stem cell cassette [101].”
From Non-Integrating Lentiviral Vectors in Clinical Applications: A Glance Through (2022) · cites this paper as [101]
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“Fibroblasts from four previously diagnosed FAP individuals were obtained from Coriell and reprogrammed using the STEMCCA system to generate three independent iPSC lines, as described [12, 18]”
From Modeling APC mutagenesis and familial adenomatous polyposis using human iPS cells (2018) · cites this paper as [18]
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Metadata
- Authors
- Sommer, C. A., Stadtfeld, M., Murphy, G. J., Hochedlinger, K., Kotton, D. N., & Mostoslavsky, G.
- Affiliations
- Boston University (US)
- Massachusetts General Hospital (US)
- Harvard University (US)
- Corresponding author
- Mostoslavsky, Gustavo
- ORCID
- Venue
- Stem Cells
- Publisher
- Oxford University Press (OUP)
- Published
- 2009-03-01
- Volume
- 27
- Issue
- 3
- Pages
- 543-549
- ISSN
- 1066-5099 (ISSN-L)
- 1549-4918
- PMC
- PMC4848035
- OpenAlex
- W2038815755
- Cited by
- 676
- References
- 24
- Citations by year
- 2026: 6 · 2025: 13 · 2024: 16 · 2023: 15 · 2022: 18 · 2021: 23 · 2020: 30 · 2019: 35 · 2018: 42 · 2017: 41 · 2016: 41 · 2015: 49 · 2014: 61 · 2013: 65 · 2012: 66
- FWCI
- 21.14
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Stem Cells (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- V Foundation for Cancer Research
- National Institutes of Health (1dp2od003266-01)
- Harvard Stem Cell Institute
- National Heart, Lung, and Blood Institute (P01HL047049)
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (R21HD060308)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Mesenchymal stem cell research
- Keywords
- induced pluripotent stem cells, lentiviral vectors, Oct4, KLF4, SOX2, c-Myc, 2A peptide, internal ribosome entry site, fibroblast reprogramming, chromosomal integration, teratoma assay, mouse chimeras, pluripotency
- MeSH terms
- Kruppel-Like Factor 4, Animals, Cells, Cultured, Genetic Vectors, Blotting, Southern, Lentivirus, Proto-Oncogene Proteins c-myc, Reverse Transcriptase Polymerase Chain Reaction, Pluripotent Stem Cells, Octamer Transcription Factor-3, Mice, Kruppel-Like Transcription Factors, SOXB1 Transcription Factors
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