Use Of siRNA To Achieve Down Regulation Of An Endogenous Gene In Combination With The Use of A Sense Construct To Achieve Expression Of A Polynucleotide

Abstract

The present invention relates to the combinatorial use of an siRNA targeted against an endogenous gene to knock out or knock down expression of the endogenous gene in a host and a delivery of a polynucleotide encoding the gene in a delivery vehicle/expression vector to the host to provide expression in the host of the protein encoded by the polynucleotide.

1 . A composition comprising a complex of an eIF5A1 siRNA targeted against the 3′ end of eIF5A1, an expression vector comprising a polynucleotide encoding a mutant eIF5A1 wherein the mutant eIF5A1 is unable to be hypusinated, and wherein the siRNA and the expression vector are complexed to polyethylenimine to form a complex. 
     
     
         2 . A composition comprising an siRNA targeted against a target gene to suppress endogenous expression of the target gene in a subject; and a polynucleotide encoding a target protein capable of being expressed in the subject in an RNAI resistant plasmid, wherein the siRNA and the plasmid are complexed to polyethylenimine to form a complex. 
     
     
         3 . The composition of  claim 1  wherein the siRNA has the sequence shown in  FIG. 25  and wherein the polynucleotide encoding the mutant eIF5A1 is eIF5A1 K50R . 
     
     
         4 . The composition of  claim 3  comprising a tissue specific promoter. 
     
     
         5 . The composition of  claim 4  comprising a B cell specific promoter. 
     
     
         6 . The composition of  claim 5  wherein the B cell promoter is B29. 
     
     
         7 . The composition of  claim 3  wherein the expression vector comprises a pCpG plasmid. 
     
     
         8 . The composition of  claim 1  wherein the eIF5A1 siRNA and the expression vector comprising the mutant eIF5A1 polynucleotide are independently complexed to polyethylenimine. 
     
     
         9 . The composition of  claim 1  wherein the eIF5A1 siRNA and the expression vector comprising the mutant eIF5A1 polynucleotide are together complexed to polyethylenimine. 
     
     
         10 . A composition comprising an eIF5A1 siRNA targeted against the 3′ end of eIF5A1 and an expression vector comprising a polynucleotide encoding a mutant eIF5A1 wherein the mutant eIF5A1 is unable to be hypusinated, and wherein the siRNA and the expression vector are delivered to a subject to treat cancer. 
     
     
         11 . The composition of  claim 10 , wherein the cancer is multiple myeloma. 
     
     
         12 . A method of treating cancer comprising administering the composition of  claim 10  to a subject. 
     
     
         13 . A method of treating cancer comprising administering the composition of  claim 1  to a subject. 
     
     
         14 . The method of  claim 12  wherein the composition is administered intravenously, intra peritoneally or intra tumorally. 
     
     
         15 . The method of  claim 13  wherein the siRNA targeted against the 3′ end of eIF5A1 and the expression vector comprising the polynucleotide encoding a mutant eIF5A1 are delivered via different routes. 
     
     
         16 . The method of  claim 12  wherein the composition is provided at a dose of about 0.15 mg/kg to about 1.5 mg/kg for twice weekly injections. 
     
     
         17 . The method of  claim 12  wherein the composition is provided at a dose of about 0.75 mg/kg to about 1.5 mg/kg for twice weekly injections.