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- Title
Gene Editing of Human Embryonic Stem Cells via an Engineered Baculoviral Vector Carrying Zinc-finger Nucleases.
- Authors
Lei, Yuning; Lee, Chi-Lin; Joo, Kye-Il; Zarzar, Jonathan; Liu, Yarong; Dai, Bingbing; Fox, Victoria; Wang, Pin
- Abstract
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.
- Subjects
STEM cell research; GENETIC engineering; RECOMBINANT baculoviruses; ZINC-finger proteins; NUCLEASES; BACULOVIRUS genetics; GENETIC vectors; REGENERATIVE medicine
- Publication
Molecular Therapy, 2011, Vol 19, Issue 5, p942
- ISSN
1525-0016
- Publication type
Article
- DOI
10.1038/mt.2011.12