Gene editing of human embryonic stem cells via an engineered baculoviral vector carrying zinc-finger nucleases
Open accessAbstract
Human embryonic stem (hES) cells are renewable cell sources that have potential applications in regenerative medicine. The development of technologies to produce permanent and site-specific genome modifications is in demand to achieve future medical implementation of hES cells. We report herein that a baculoviral vector (BV) system carrying zinc-finger nucleases (ZFNs) can successfully modify the hES cell genome. BV-mediated transient expression of ZFNs specifically disrupted the CCR5 locus in transduced cells and the modified cells exhibited resistance to HIV-1 transduction. To convert the BV to a gene targeting vector, a DNA donor template and ZFNs were incorporated into the vector. These hybrid vectors yielded permanent site-specific gene addition in both immortalized human cell lines (10%) and hES cells (5%). Modified hES cells were both karyotypically normal and pluripotent. These results suggest that this baculoviral delivery system can be engineered for site-specific genetic manipulation in hES cells.
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Baculoviral vector has been used to transduce hES cells to achieve a 5% efficacy of CCR5 disruption.
“In addition baculoviral vector has also been used to transduce hES cells to achieve a 5% efficacy of CCR5 disruption [58].”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [58]
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References 46 · Cited by 42
- Authors
- Lei, Y., Lee, C.-L., Joo, K.-I., Zarzar, J., Liu, Y., Dai, B., Fox, V., & Wang, P.
- Affiliations
- University of Southern California (US)
- Corresponding author
- Fox, Victoria
- Wang, Pin
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2011-05
- Volume
- 19
- Issue
- 5
- Pages
- 942-950
- ISSN
- 1525-0016 (ISSN-L)
- PMC
- PMC3098635
- OpenAlex
- W2115763396
- Cited by
- 42
- References
- 46
- Citations by year
- 2025: 3 · 2024: 2 · 2021: 1 · 2019: 1 · 2018: 1 · 2017: 1 · 2016: 7 · 2015: 4 · 2014: 6 · 2013: 9 · 2012: 3
- FWCI
- 2.86
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- California Institute for Regenerative Medicine
- California Institute of Regenerative Medicine
- National Institute of Allergy and Infectious Diseases (R01AI068978)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- human embryonic stem cells, zinc finger nucleases, HIV-1 resistance, gene targeting, transgene delivery, pluripotency
- MeSH terms
- Cell Line, Deoxyribonucleases, Flow Cytometry, Genetic Vectors, Humans, Mutation, Transduction, Genetic, HIV-1, Zinc Fingers, Nucleopolyhedroviruses, Gene Targeting, Transgenes, Receptors, CCR5, Reverse Transcriptase Polymerase Chain Reaction, Embryonic Stem Cells
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