Transcriptome transfer produces cellular phenotype conversion
The present invention includes methods for effecting phenotype conversion in a cell by transfecting the cell with phenotype-converting nucleic acid. Expression of the nucleic acids results in a phenotype conversion in the transfected cell. Preferably the phenotype-converting nucleic acid is a transcriptome, and more preferably an mRNA transcriptome.
1. A method of effecting phenotype conversion in a cell, said method comprising introducing phenotype-converting nucleic acid of a first cell having a phenotype into a second cell having a phenotype, wherein the phenotype-converting nucleic acid comprises at least one RNA, wherein the phenotype of said first cell is different from the phenotype of said second cell, wherein the second cell is pre-treated with a transcription inhibitor before it is transfected, wherein said transfected phenotype-converting nucleic acid causes the phenotype of said second cell to change to the phenotype of said first cell, and wherein said phenotype-converting nucleic acid is locally administered to said second cell in vivo.
2. The method of claim 1 , wherein said phenotype-converting nucleic acid is an mRNA transcriptome.
3. The method of claim 1 , wherein said phenotype of said first cell differs from said phenotype of said second cell by one or more of: differentiation degree, disease state, exposure to a toxin, exposure to a pathogen, and exposure to a candidate therapeutic.
4. The method of claim 2 , wherein said mRNA transcriptome comprises mRNA transcripts having an average size between about 1 kb to about 5 kb.
5. The method of claim 2 , further comprising introducing said second cell at least a second time with said first cell mRNA transcriptome.
6. The method of claim 1 , wherein said first cell is a cardiomyocyte.
7. The method of claim 1 , wherein said second cell is a fibroblast.
8. The method of claim 1 , wherein said transcription inhibitor is 5,6-Dichloro-1-β-D-ribofuranosylbenzimidazole (DRB).
9. The method of claim 1 , wherein said first cell and said second cell are each selected from the group consisting of a eukaryotic cell and a prokaryotic cell.
10. The method of claim 9 , wherein said eukaryotic cell is a non-mammalian cell.
11. The method of claim 9 , wherein said eukaryotic cell is a mammalian cell.
12. The method of claim 9 , wherein said eukaryotic cell is a human cell.
13. The method of claim 2 , wherein said phenotype-converting nucleic acid further comprises one or more exogenous nucleic acids selected from the group consisting of mRNA, siRNA, miRNA, hnRNA, tRNA, non-coding RNA and combinations thereof.
14. The method of claim 1 , wherein said phenotype conversion comprises a change in one or more of gene expression, protein expression, immunological markers, morphology, physiology, synthesis of bioproducts, and membrane lipid composition.
15. The method of claim 14 , wherein phenotype conversion comprises a change in expression of at least 100 genes.
16. The method of claim 14 , wherein phenotype conversion comprises up-regulation of genes associated with chromosomal remodeling.
17. The method of claim 14 , wherein at least about 5% of differentially expressed genes in said second cell change expression to a level observed for said first cell.
18. The method of claim 1 , wherein said phenotype conversion persists for at least 2 weeks.
19. The method of claim 18 , wherein said phenotype conversion persists for the lifetime of the cell.