title: [Inhibition of HIV-1 in vitro by combination of vpr and tat specific short hairpin RNA via lentiviral vectors].
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.
authors.affiliation: The College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100022, China. hzhaoyanzhe@yahoo.com.cn
authors.forename: Yan-Zhe
authors.initials: YZ
authors.lastname: Hao
authors.forename: Zhi-Ping
authors.initials: ZP
authors.lastname: Teng
authors.forename: Yi-Shu
authors.initials: YS
authors.lastname: Yang
authors.forename: Xiao-Na
authors.initials: XN
authors.lastname: Sun
authors.forename: Jing
authors.initials: J
authors.lastname: Ma
authors.forename: Xiao-Hua
authors.initials: XH
authors.lastname: Jin
authors.forename: Yi
authors.initials: Y
authors.lastname: Zeng
mesh_terms: D002460:Cell Line; D015536:Down-Regulation; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D015658:HIV Infections / Q000628:therapy / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000378:metabolism; D054325:vpr Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D054322:tat Gene Products, Human Immunodeficiency Virus; D054325:vpr Gene Products, Human Immunodeficiency Virus; C514912:vpr protein, Human immunodeficiency virus 1