Delivery of packaged RNA to mammalian cells
AssigneeThe United States of America, 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.
Claims8
A recombinant alphavirus particle, comprising:
- a. an alphavirus replicon vector construct generated from a Sindbis virus or Venezuelan equine encephalitis virus which directs the expression of a heterologous nucleic acid molecule, wherein said alphavirus replicon vector construct comprises (i) a 5′ sequence which initiates transcription of alphavirus RNA, (ii) a Rous sarcoma virus packaging signal, (iii) a nucleotide sequence encoding Sindbis virus or Venezuelan equine encephalitis virus nonstructural proteins NSP1, NSP2 NSP3 and NSP4, (iv) a viral junction region promoter which directs the expression of the heterologous nucleic acid sequence, and (v) an RNA polymerase recognition sequence, wherein said heterologous nucleic acid sequence replaces an alphavirus structural protein gene;
- b. a Rous sarcoma virus capsid protein, and
- c. an envelope glycoprotein or fusogenic protein heterologous to the alphavirus of said alphavirus construct and capable of binding to a host eukaryotic cell.
Dependencies are inferred from the claim text.