Gene editing of human embryonic stem cells via an engineered baculoviral vector carrying zinc-finger nucleases

Yuning Lei, Chi Lin Lee, Kye Il Joo, Jonathan Zarzar, Yarong Liu, Bingbing Dai, Victoria Fox, Pin Wang

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

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 languageEnglish
Pages (from-to)942-950
Number of pages9
JournalMolecular Therapy
Volume19
Issue number5
DOIs
StatePublished - 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.

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