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US 10,801,041 B2Granted
T7 alpha viral vector system
AssigneeOrbis Health Solutions LLC
InventorThomas E. Wagner
Life of this patent
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Abstract
The present invention relates generally to a gene expression system utilizing an alphavirus replicon and T7 promoter. The system is capable of expressing proteins in the cell cytoplasm without integrating the gene of interest into the genome of a host cell. The invention has a wide range of applications such as producing induced pluripotent cells and vaccines against pathogens and cancers.
Claims20
A gene expression system comprising
- (a) a vector that comprises in a 5′ to 3′ orientation a bacteriophage promoter, a first capping sequence, an alphavirus replicon operatively linked to the bacteriophage promoter, a subgenomic promoter, a second capping sequence, and a gene of interest, and
- (b) a protein that comprises an exogenous RNA polymerase which transcribes the alphavirus replicon, wherein the exogenous RNA polymerase is co-delivered with the alphavirus vector into a cell,
- and wherein the gene of interest is expressed in the cytoplasm of the cell.
A method for producing an induced pluripotent stem cell, comprising
- (a) introducing into a mammalian somatic cell a gene expression system comprising a vector, wherein the vector comprises a sequence that encodes one or more mammal-derived reprogramming factors, and an exogenous RNA polymerase co-delivered with the vector, and
- (b) culturing the transduced somatic cell in the presence of a cytokine on a fibroblast feeder layer or an extracellular matrix under a condition for maintaining pluripotency and self-renewal, wherein the cytokine is basic fibroblast growth factor (bFGF) or stem cell factor (SCF).
Dependencies are inferred from the claim text.