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.
| Original language | English |
|---|---|
| Pages (from-to) | 942-950 |
| Number of pages | 9 |
| Journal | Molecular Therapy |
| Volume | 19 |
| Issue number | 5 |
| DOIs | |
| State | Published - May 2011 |
Bibliographical note
Funding Information:This work was supported by grant from the California Institute for Regenerative Medicine (RT1-01028-1). We thank Steven Froelich and April Tai for critical reading of this manuscript.