World J Stem Cells. 2013 October 26; 5(4): 98-105.
Published online 2013 October 26. doi: 10.4252/wjsc.v5.i4.98.
Advances in homology directed genetic engineering of human pluripotent and adult stem cells
Kalpith Ramamoorthi, Donald Curtis and Prashanth Asuri.
Kalpith Ramamoorthi, Donald Curtis, Prashanth Asuri, Department of Bioengineering, Santa Clara University, Santa Clara, CA 95053, United States
Author contributions: All authors have contributed to this article.
Correspondence to: Prashanth Asuri, PhD, Department of Bioengineering, Santa Clara University, 500 El Camino Real, Santa Clara, CA 95053, United States. asurip@scu.edu
Telephone: +1-408-5513005 Fax: +1-408-5545474
Received August 15, 2013; Revised September 6, 2013; Accepted September 14, 2013;
Abstract
The ability to introduce precise genomic modifications in human cells has profound implications for both basic and applied research in stem cells, ranging from identification of genes regulating stem cell self-renewal and multilineage differentiation to therapeutic gene correction and creation of in vitro models of human diseases. However, the overall efficiency of this process is challenged by several factors including inefficient gene delivery into stem cells and low rates of homology directed site-specific targeting. Recent studies report the development of novel techniques to improve gene targeting efficiencies in human stem cells; these methods include molecular engineering of viral vectors to efficiently deliver episomal genetic sequences that can participate in homology directed targeting, as well as the design of synthetic proteins that can introduce double-stranded breaks in DNA to initiate such recombination events. This review focuses on the potential of these new technologies to precisely alter the human stem cell genome and also highlights the possibilities offered by the combination of these complementary strategies.
Keywords: Human stem cells, Genetic engineering, Engineered viruses, Synthetic restriction endonucleases