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.
Claims7
A replication defective virus like particle (VLP) comprising:
- a. an alphavirus replicon isolated from a Sindbis virus or Venezuelan equine encephalitis (VEE) virus comprising a recombinant polynucleotide comprising:
- (i) a 5′ sequence which initiates transcription of alphavirus RNA, (ii) a Rous sarcoma virus (RSV) packaging signal, (iii) a nucleotide sequence encoding Sindbis virus or Venezuelan equine encephalitis virus nonstructural proteins NSP1, NSP2 NSP3 and NSP4, (iv) one or more subgenomic mRNA promoter (sgp) for Sindbis or VEE-encoded RNA-dependent RNA polymerase, wherein said sgp is linked to a heterologous nucleic acid sequence, (v) a 3′ untranslated region (URT) and (vi) a polyA tail, wherein said heterologous nucleic acid sequence replaces the alphavirus structural protein gene;
- b. a retroviral gag protein encoded by a polynucleotide comprising a genomic sequence of the RSV,
- c. a fusogenic envelope protein,
- wherein the VLP does not comprise or express a retroviral pol gene, and wherein the VLP is not cytopathic to a eukaryotic cell.
Dependencies are inferred from the claim text.