Molecular clones with mutated HIV gag/pol, SIV gag and SIV env genes
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorGeorge N. Pavlakis
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Abstract
Nucleic acid constructs containing HIV-1 gag/pol and SIV gag or SIV env genes which have been mutated to remove or reduce inhibitory/instability sequences are disclosed. Viral particles and host cells containing these constructs and/or viral particles are also disclosed. The exemplified constructs and viral particles of the invention may be useful in gene therapy for numerous disorders, including HIV infection, or as a vaccine for HIV-1 immunotherapy and immunoprophylaxis.
Claims22
A nucleic acid construct comprising a HIV-1 gag/pol gene having the coding sequence of the gag/pol gene set forth in FIG. 1 (SEQUENCE ID NO:
- 1).
A nucleic acid construct comprising a HIV-1 pol gene having the coding sequence of the pol gene set forth in FIG. 2 (SEQUENCE ID NO:
- 3).
A nucleic acid construct comprising an HIV or SIV 5′ LTR, a packaging signal, a gag/pol gene comprising the sequence set forth in FIG. 1 (SEQUENCE ID NO:
- 1), a 5′ splice site, a 3′ splice site, an env gene, a tat gene, a functional RNA transport element and a 3′ HIV or SIV LTR, said nucleic acid construct being able to produce functional Gag. Pol and Env virion components.
A lentiviral expression system comprising the following:
- (a) a packaging vector comprising a HIV-1 gag/pol gene having the nucleotide sequence set forth in FIG. 1 (SEQUENCE ID NO:
- 1);
- (b) a transfer vector;
- and
- (c) an envelope encoding vector.
A process for making a lentiviral particle comprising expressing, in a host cell, HIV Gag and HIV Pol from a vector comprising the nucleotide sequences encoding HIV Gag and HIV Pol set forth in FIG. 1 (SEQUENCE ID NO:
- 1) and expressing a gene encoding an envelope protein by growing the host cell under conditions suitable to cause expression of HIV Gag, HIV Pol and envelope protein so that a lentiviral particle is formed.
A lentiviral expression system which is capable of functioning in the absence of Rev, Tat, and any viral RNA transport element comprising the following:
- (a) a packaging vector comprising a HIV-1 gag/pol gene which is capable of functioning in the absence of Rev, Tat, and any viral RNA transport element;
- (b) a transfer vector;
- and
- (c) an envelope encoding vector
- wherein the HIV-1 gag/pol gene has the coding sequence of the HIV-1 gag/pol gene set forth in FIG. 1 (SEQUENCE ID NO:
- 1).
A process for making a lentiviral particle in the absence of Rev, Tat, or any viral RNA transport element comprising expressing HIV Gag and HIV Pol in a host cell from a HIV-1 gag/pol gene which is capable of functioning in the absence of Rev, Tat, and any viral RNA transport element and expressing an Envelope protein from a envelope encoding gene whose expression is independent of Rev, Tat, or any viral RNA transport element wherein the HIV-1 gag/pol gene has the coding sequence of the HIV-1 gag/pol gene set forth in FIG. 1 (SEQUENCE ID NO:
- 1).
Dependencies are inferred from the claim text.