Transcriptome transfer produces cellular phenotype conversion

Patent metadata

Field	Value
Publication number	US-9845480-B2
Application number	US-201514880737-A
Country	US
Kind code	B2
Filing date	Oct 12, 2015
Priority date	Dec 13, 2005
Publication date	Dec 19, 2017
Grant date	Dec 19, 2017

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Univ Pennsylvania

Inventors

• Eberwine James
• Sul Jai-Yoon
• Kim Tae Kyung
• Patel Vickas

Classifications

• C12N2506/08

  from cells of the nervous system · CPC title

• A61K31/713

  Double-stranded nucleic acids or oligonucleotides · CPC title

• A61K45/06

  Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca · CPC title

• C12N2506/00

  Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells · CPC title

• C12N15/85 Primary

  for animal cells · CPC title