Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 182, Issue 1-2, Pages 9-17Publisher
ELSEVIER
DOI: 10.1016/j.jviromet.2012.02.007
Keywords
RNAi; Cell sorting; Human immunodeficiency virus; Animal model; Lentivirus
Funding
- National Natural Science Foundation of China [81172895, 81172885]
- National Natural Science Foundation [30925030]
- Science and Technology Major Project of Fujian Province of China [2010Y4008, 2011J06015]
- International Science and Technology Collaborative Program of China [2010DFB30100]
Ask authors/readers for more resources
Gene therapy offers a potentially an effective treatment for many human diseases, including HIV/AIDS. One of the most studied gene delivery systems is the use of lentivirus based vectors, which can deliver genes into both dividing and nondividing cells. However, low infection efficiency represents an obstacle for proper evaluation of their biological function. In this study, a recombinant lentiviral vector which expressed short hairpin RNAs (shRNAs) targeted against the HIV-1 vif/pol was transduced into various cells. An MHC class I molecule, H-2K(k) was used as a marker to accumulate the virally transduced cells through immunomagnetic sorting. In vitro testing of transduced cells showed 85% suppression of HIV in post-sorted PBMCs compared to 30% in pre-sorted PBMCs. In additional, using a mouse xenotrans-plantation model with the same treatment protocol for cell enrichment, a > 95% decrease in HIV activity in post-sorted cells was achieved, as compared to nearly none in the pre-sorted cells. These studies offer a practical method to accumulate virally transduced cells, which can be applied to evaluate the performance of various shRNAs constructs. (C) 2012 Elsevier BM. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available