Expression cassette and vector for transient or stable expression of exogenous molecules

Abstract

The disclosure provides an expression cassette and a vector comprising the cassette for expression of a polynucleotide. The expression cassette includes a promoter/enhancer, an intervening region, and a polyadenylation signal domain. Expression systems and methods of using the expression cassette and vector are also provided.

1 - 13 . (canceled) 
     
     
         14 . An expression cassette comprising:
 a human CMV immediate early 1 (hCMV IE1) promoter/enhancer region;   a polynucleotide of interest; and   a polyA signal domain comprising SEQ ID NO: 19.   
     
     
         15 . The expression cassette of  claim 14 , further comprising a variable length intervening sequence (VLIVS) comprising a splice donor and splice acceptor site. 
     
     
         16 . The expression cassette of  claim 15 , wherein the VLIVS comprises an intron A of a hCMV IE1 gene that has a deletion between the splice acceptor and splice donor of the intron A. 
     
     
         17 . The expression cassette of  claim 14 , further comprising an additional promoter/enhancer element or regulatory region. 
     
     
         18 . The expression cassette of  claim 17 , wherein the promoter/enhancer element is an SV40 promoter/enhancer element. 
     
     
         19 . The expression cassette of  claim 17 , wherein the regulatory region is an SV40 polyadenylation region. 
     
     
         20 . The expression cassette of  claim 14 , further comprising a selectable marker. 
     
     
         21 . The expression cassette of  claim 20 , wherein the selectable marker is selected from the group consisting of dihydrofolate reductase, GFP, neomycin, Hygro, Zeocin™, herpes simplex virus thymidine kinase, hypoxanthine-guanine phosphoribosyltransferase, adenine phosphoribosyltransferase, puromycin N-acetyl transferase or adenosine deaminase. 
     
     
         22 . The expression cassette of  claim 21 , wherein the selectable marker is dihydrofolate reductase. 
     
     
         23 . The expression cassette of  claim 14 , wherein the polynucleotide of interest encodes a therapeutic agent. 
     
     
         24 . The expression cassette of  claim 14 , wherein the polynucleotide of interest is in the antisense orientation with respect to the promoter. 
     
     
         25 . The expression cassette of  claim 14 , wherein the polynucleotide of interest is a heterologous polynucleotide. 
     
     
         26 . The expression cassette of  claim 14 , wherein the polynucleotide of interest further includes one or more transcriptional control elements. 
     
     
         27 . The expression cassette of  claim 26 , wherein the transcriptional control elements are selected from the group consisting of transcriptional stop signals, polyadenylation domains and downstream enhancer elements. 
     
     
         28 . The expression cassette of  claim 14 , wherein the polynucleotide of interest encodes a polypeptide of diagnostic or therapeutic use. 
     
     
         29 . The expression cassette of  claim 14 , wherein the polynucleotide of interest encodes an antigenic polypeptide for use as a vaccine. 
     
     
         30 . The expression cassette of  claim 14  in the form of a naked nucleic acid construct. 
     
     
         31 . An expression vector comprising an expression cassette of  claim 14 . 
     
     
         32 . A host cell comprising an expression vector of  claim 31 . 
     
     
         33 . A method for producing a polypeptide comprising propagation of a host cell of  claim 32  and expression of the polynucleotide of interest. 
     
     
         34 . A method of delivering a therapeutic agent to an animal in need of treatment comprising propagation of a host cell of  claim 33  and expression of the polynucleotide of interest. 
     
     
         35 . A method of gene therapy to an animal in need of treatment comprising propagation of a host cell of  claim 33  and expression of the polynucleotide of interest. 
     
     
         36 . A polynucleotide that specifically hybridizes to a polynucleotide sequence as set forth in SEQ ID NO: 1 from about nucleotide 1 to about 1867 or a fragment thereof.