Delivery of packaged rna to mammalian cells
AssigneeThe USA, as represented by the Secretary, Dept. of Health and Human Services
InventorsStanislaw J. Kaczmarczyk, Deb K. Chatterjee
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Abstract
Described herein are compositions relating to alphavirus-based virus-like particles (VLPs) and methods for making and using the described VLPs. The described compositions include VLPs and vectors and cells used to produce the VLPs. Also included are related methods to produce the VLPs, to transduce cells using the VLPs, and to produce a protein or polynucleotide of interest in a target cell using the VLPs. Also described are alphavirus-based replicons that allow for expression of proteins or polynucleotides of interest in a target cell without a cytopathic effect.
Claims9
A virus like particle (VLP) consisting of:
- a. an alphavirus replicon isolated from Sindbis virus or Venezuelan equine encephalitis virus comprising a recombinant polynucleotide comprising a sequence encoding Sindbis virus or Venezuelan equine encephalitis virus nonstructural proteins NSP1, NSP2, NSP3, and NSP4;
- and a retroviral packaging signal isolated from Rous sarcoma virus,
- b. a retroviral gag protein encoded by a polynucleotide comprising a genomic sequence of a Rous sarcoma virus, and
- c. a fusogenic envelope protein,
- wherein the VLP does not contain an alphavirus structural protein gene.
A method of producing a VLP comprising:
- a. co-transforming a eukaryotic cell with:
- i. a first vector comprising a polynucleotide sequence encoding the alphavirus replicon, wherein the alphavirus replicon includes the polynucleotide of interest,
- ii. a second vector comprising a polynucleotide sequence encoding the retroviral Rous gag protein, and
- iii. a third vector comprising a polynucleotide sequence encoding the fusogenic envelope protein;
- b. culturing the co-transformed cell under conditions suitable to cause each vector to produce its encoded product, thereby producing the VLP;
- and
- c. isolating the VLP from the cell;
- wherein neither the vectors nor the cell contain any alphavirus structural protein genes.
Dependencies are inferred from the claim text.