Inhibition of HIV-1 in vitro by combination of vpr and tat specific short hairpin RNA via lentiviral vectors.
- Author:
Yan-Zhe HAO
1
;
Zhi-Ping TENG
;
Yi-Shu YANG
;
Xiao-Na SUN
;
Jing MA
;
Xiao-Hua JIN
;
Yi ZENG
Author Information
1. The College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100022, China. hzhaoyanzhe@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Cell Line;
Down-Regulation;
Genetic Therapy;
Genetic Vectors;
genetics;
metabolism;
HIV Infections;
therapy;
virology;
HIV-1;
genetics;
metabolism;
Humans;
Lentivirus;
genetics;
metabolism;
RNA Interference;
RNA, Small Interfering;
genetics;
metabolism;
therapeutic use;
tat Gene Products, Human Immunodeficiency Virus;
genetics;
metabolism;
vpr Gene Products, Human Immunodeficiency Virus;
genetics;
metabolism
- From:
Chinese Journal of Virology
2013;29(2):126-131
- CountryChina
- Language:Chinese
-
Abstract:
Gene therapy offers the promise of curing the HIV-infected patients. Specific, potent, and sustained short hairpin RNA (shRNA)-mediated gene silencing is crucial for the successful application of RNA interference technology to therapeutic interventions. To reduce the probability of viral escape mutants, in this study, we constructed lentiviral vector containing vpr and tat shRNA, respectively, furthermore the bispecific lentiviral vector harboring vpr and tat shRNA expression cassettes from U6 promotor and H1 promotor was cotransfected with recombinant plasmid expressing the vpr and tat gene. The result showed that the bispecific lentiviral vector plvx-vpr-tatshRNA could inhibit the vpr and tat effectively,with ratios of 89.20% and 62.00% respectively. When cotransfected with pNL4-3 in 293T cell, plvx-vpr-tatshRNA showed higher efficacy in down regulating the HIV NL4-3 packaging production than the plvx-vprshRNA or plvx-tatshRNA individually. MT4 cell clones transduced with recombinant lentiviral vectors were screened and challenged with HIV NL4-3. P24 ELISA test showed that MT4 transduced with the combinational lentiviral vector could inhibit virus replication efficiently.
