Inhibition of human immunodeficiency virus type 1 (HIV-1) replication by HIV-1-based lentivirus vectors expressing transdominant Rev.
Retrovirus vectors expressing transdominant-negative mutants of Rev (TdRev) inhibit human immunodeficiency virus type 1 (HIV-1) replication by preventing the nuclear export of unspliced viral transcripts, thus inhibiting the synthesis of Gag-Pol, Env, and genomic RNA. The use of HIV-1-based vectors to express TdRev would have the advantage of allowing access to nondividing hematopoietic cells. It would also provide additional levels of protection by sequestering the viral regulatory proteins Tat and Rev, competing for encapsidation into wild-type virions, and inhibiting reverse transcription. Here we describe HIV-1-based vectors that express TdRev. These vectors contain mutations in the splicing signals or replacement of the Rev-responsive element by the simian retrovirus type 1 constitutive transport element, making them less sensitive to the inhibitory effects of TdRev. In addition, overexpression of Rev and the use of an HIV-1 helper plasmid that drives high levels of Gag-Pol synthesis were used to transiently overcome the inhibition by TdRev of the synthesis of Gag-Pol during vector production. SupT1 cells transduced with these vectors were more resistant to HIV-1 replication than cells transduced with Moloney murine leukemia virus-based vectors expressing TdRev. Furthermore, we show that these vectors can be mobilized by the wild-type virus, reducing the infectivity of virions escaping inhibition and conferring protection against HIV-1 replication to previously untransduced cells.
Clinical Gene Therapy Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892-1851, USA.
M R
MR
Mautino
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N
Keiser
R A
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Morgan
D015139:Blotting, Southern; D002460:Cell Line; D015685:Fusion Proteins, gag-pol / Q000096:biosynthesis; D015870:Gene Expression; D015694:Gene Products, rev / Q000235:genetics / Q000502:physiology*; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D016655:HIV Core Protein p24 / Q000096:biosynthesis; D015497:HIV-1 / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008979:Moloney murine leukemia virus / Q000235:genetics; D011533:Proviruses / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism / Q000821:virology; D014161:Transduction, Genetic; D019065:Virus Assembly; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus
D015685:Fusion Proteins, gag-pol; D015694:Gene Products, rev; D016655:HIV Core Protein p24; D054321:rev Gene Products, Human Immunodeficiency Virus