Autogene nucleic acids encoding a secretable RNA polymerase

1. A secretable RNA polymerase expression cassette comprising:

(a) a eukaryotic promoter and a first RNA polymerase promoter operably linked to a nucleic acid encoding a secretable RNA polymerase (sRNAP) comprising a RNA polymerase and a secretion domain; and

(b) a second RNA polymerase promoter operably linked to a nucleic acid encoding a product of interest, wherein the sRNAP enters the cytoplasm of a cell and carries out template dependent synthesis of RNA.

2. The expression cassette of claim 1 , wherein said RNA polymerase is a non-host RNA polymerase.

3. The expression cassette of claim 1 , wherein said RNA polymerase is a member selected from the group consisting of: a phagemid RNA polymerase, a prokaryotic RNA polymerase, an archaebacterial RNA polymerase, a plant RNA polymerase, a fungal RNA polymerase, a eukaryotic RNA polymerase, a viral RNA polymerase, mitochondrial RNA polymerase, and a chloroplast RNA polymerase.

4. The expression cassette of claim 3 , wherein said phagemid RNA polymerase is a member selected from the group consisting of: a SP6 RNA Polymerase, a T7 RNA Polymerase, a K11 RNA Polymerase, and a T3 RNA Polymerase.

5. The expression cassette of claim 1 , wherein said product of interest is a member selected from the group consisting of: a protein, a nucleic acid, an antisense nucleic acid, ribozymes, tRNA, snRNA, and an antigen.

6. The expression cassette of claim 1 , wherein said product of interest encodes a protein selected from the group consisting of:

a herpes simplex virus thymidine kinase (HSV-TK), a cytosine deaminase, a xanthine-guaninephosphoribosyl transferase, a purine nucleoside phosphorylase, a carboxylesterase, a deoxycytidine kinase, a nitroreductase, a thyrnidine phosphorylase, and a cytochrome P450 2B1.

7. The expression cassette of claim 1 , wherein said product of interest encodes a protein selected from the group consisting of: p53, a retinoblastoma susceptibility gene product (RB1), a calmodulin-dependent serine threonine (DAP kinase), p16, a protein from an alternative reading frame of p16 (ARF), an adenomatous polyposis coli gene product (APC), neurofibromin, a phosphatase and tensin homologue deleted from chromosome 10 (PTEN), a Wilms= tumor gene product (WT1), a neurofibromatosis type 1 gene product (NF1), and a von Hippel-Lindau tumor suppressor gene product (VHL).

8. The expression cassette of claim 1 , wherein said product of interest encodes a protein selected from the group consisting of: angiostatin, endostatin, and vascular endothelial growth factor (VEGF)-R2.

9. The expression cassette of claim 1 , wherein said product of interest encodes apoptin.

10. The expression cassette of claim 1 , wherein said product of interest encodes a protein selected from the group consisting of: IL-2, IL-3, IL-4, IL-6, IL-7, IL-10, IL-12, IL-15, IFN-α, IFN-β, IFN-γ, TNF-α, GM-CSF, G-CSF, and Fms-like tyrosine kinase 3 (Flt3)-Ligand.

11. The expression cassette of claim 1 , wherein said product of interest is a protein selected from the group consisting of: a restriction endonuclease, a single chain antibody, a peptide hormone, erythropoietin (EPO), and single-chain insulin.

12. The expression cassette of claim 1 , wherein said eukaryotic promoter is a member selected from a group consisting of: a cytomegalovirus promoter, a von Willebrand factor (vWf) promoter, a Clara cell secretory protein (CCSP/UG) promoter, an osteoblast-specific osteocalcin promoter, an albumin promoter, a muscle creatine kinase (MCK) promoter, a mucin-1 (Muc-1) promoter, a carcinoembryonic antigen (CEA) promoter, a prostate specific antigen (PSA) promoter, an epidermal growth factor receptor (HER-2) promoter, a Myc promoter, a L-plastin promoter, an alpha-fetoprotein (AFP) promoter, a hypoxia-responsive element (HIRE) promoter, an early growth response (egr-1) promoter, a multidrug resistance (mdr-1) promoter, a heat shock protein 70 (hsp70) promoter, a tetracycline induced promoter, a simian virus 40 (SV40) promoter, an alcohol dehydrogenase (ADH1) promoter, a GAL4 promoter, a LexA promoter, a rous sarcoma virus (RSV) promoter, a human imniunodeficiency virus (HIV) promoter, and a simian foamy virus (SFV) promoter.

13. A lipid-nucleic acid composition comprising:

a nucleic acid-lipid particle comprising a lipid portion and a nucleic acid portion,

wherein said nucleic acid portion comprises a secretable RNA polymerase expression cassette comprising:

(a) a eukaryotic promoter and a first RNA polymerase promoter operably linked to a nucleic acid encoding a secretable RNA polymerase comprising a RNA polymerase and a secretion domain; and

(b) a second RNA polymerase promoter operably linked to a nucleic acid encoding a product of interest.

14. The nucleic acid of claim 13 , wherein a RNA polymerase promoter is operably linked to said nucleic acid encoding a secretable RNA polymerase.

15. The lipid-nucleic acid composition of claim 13 , wherein said nucleic acid-lipid particle is a serum-stable nucleic acid-lipid particle comprising a nucleic acid fuily encapsulated within said lipid portion.

16. The lipid-nucleic acid composition of claim 13 , wherein said lipid portion comprises a protonatable lipid having a pKa in a range of about 4 to about 11.

17. The lipid-nucleic acid composition of claim 13 , wherein said lipid portion comprises a cationic lipid.

18. The lipid-nucleic acid composition of claim 17 , wherein said cationic lipid is a member selected from the group consisting of DODAC, DODAP, DODMA, DOTAP, DOTMA, DC-Chol, DMRIE, and DSDAC.

19. The expression cassette of claim 1 , wherein said secretion domain is an importation competent signal peptide.

20. The expression cassette of claim 1 , wherein said secretion domain is a protein transduction domain.

21. The expression cassette of claim 20 , wherein said protein transduction domain is a TAT polypeptide.

22. The expression cassette of claim 21 , wherein said TAT polypeptide comprises the amino acid sequence X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -Arg-X 10 , wherein

X 1 is Tyr or Ala;

X 2 is Gly or Ala;

X 3 , X 6 , X 7 , X 9 and X 10 are Arg or Ala; and

X 8 is Gln, Arg or Ala.

23. The expression cassette of claim 22 , wherein said TAT polypeptide comprises SEQ ID NO:1.

24. The lipid-nucleic acid composition of claim 13 , wherein said secretion domain is an importation competent signal peptide.

25. The lipid-nucleic acid composition of claim 13 , wherein said secretion domain is a protein transduction domain.

26. The lipid-nucleic acid composition of claim 25 , wherein said protein transduction domain is a TAT polypeptide.

27. The lipid-nucleic acid composition of claim 26 , wherein said TAT polypeptide comprises the amino acid sequence X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -Arg-X 10 , wherein

X 1 is Tyr or Ala;

X 2 is Gly or Ala;

X 3 , X 6 , X 7 , X 9 and X 10 are Arg or Ala; and

X 8 is Gln, Arg or Ala.

28. The lipid-nucleic acid composition of claim 27 , wherein said TAT polypeptide comprises SEQ ID NO:1.

29. The expression cassette of claim 1 , wherein the RNA polymerase is a T7 RNA polymerase.

30. The lipid-nucleic acid composition of claim 13 , wherein the RNA polymerase is a T7 RNA polymerase.