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 transferring a phenotype of a first cell to a second cell, said method comprising
transfecting a second cell of a second cell type with an mRNA transcriptome of a first cell of a first cell type a first time, wherein the first cell type is different from the second cell type, wherein the second cell is transfected in vitro or ex vivo;
transfecting the second cell with an mRNA transcriptome of the first cell at least a second time wherein the second cell is transfected in vitro or ex vivo;
thereby initiating a change in physiology and morphology of the second cell, wherein the change in physiology and morphology yields a phenotype of the second cell that is indicative of the first cell
wherein the first cell and second cell are selected from the group consisting of:
a) wherein the second cell is a fibroblast, and wherein the first cell is selected from the group consisting of an astrocyte, a cardiomyocyte, and a stem cell;
b) wherein the second cell is a neuron, and wherein the first cell is selected from the group consisting of an astrocyte and a cardiomyocyte; and
c) wherein the second cell is an astrocyte and wherein the first cell is a cardiomyocyte.
2. The method of claim 1 , wherein said first cell type differs from said second cell type by one or more of: tissue type, differentiation degree, disease state, response to exposure to a toxin, response to exposure to a pathogen, and response to exposure to a candidate therapeutic.
3. The method of claim 1 , wherein said mRNA transcriptome comprises mRNA transcripts having an average size between about 1 kb to about 5 kb.
4. The method of claim 1 , wherein said transfecting step comprises irradiating said second cell with a laser, wherein said cell is bathed in a fluid comprising said first cell mRNA transcriptome.
5. The method of claim 4 , wherein said irradiating step comprises 2 to 25 laser excitation pulses, wherein said laser is directed to different sites on said second cell for each laser excitation pulse.
6. The method of claim 1 , wherein said second cell is contacted with an exogenous transcription inhibition agent prior to said transfecting step.
7. The method of claim 1 , wherein said second cell is not substantially contacted with an exogenous transcription inhibition agent before, during or after said transfecting step.
8. The method of claim 1 , wherein said first cell and said second cell are each a eukaryotic cell.
9. The method of claim 8 , wherein said eukaryotic cell is a non-mammalian cell.
10. The method of claim 8 , wherein said eukaryotic cell is a mammalian cell.
11. The method of claim 10 , wherein said eukaryotic cell is a human cell.
12. The method of claim 1 , wherein the method further comprises transfecting the second cell with one or more nucleic acids of the first cell, wherein the one or more nucleic acids comprise one or more exogenous nucleic acids selected from the group consisting of siRNA, miRNA, hnRNA, tRNA, non-coding RNA and combinations thereof.
13. The method of claim 1 , wherein the method causes the second cell to exhibit a change in one or more of gene expression, protein expression, immunological markers, synthesis of bioproducts, and membrane lipid composition.
14. The method of claim 13 , wherein the method causes the second cell to exhibit a change in expression of at least 100 genes.
15. The method of claim 13 , wherein the method causes the second cell to exhibit up-regulation of genes associated with chromosomal remodeling.
16. The method of claim 13 , wherein at least about 5% of differentially expressed genes in said second cell change expression to a level observed for said first cell.
17. The method of claim 1 , wherein the change in physiology and morphology of the second cell-persists for at least 2 weeks.
18. The method of claim 17 , wherein the change in physiology and morphology of the second cell-persists for the lifetime of the cell.
19. The method of claim 1 , wherein the second cell is a hair cell that responds to a first range of sound frequencies.
20. The method of claim 1 , wherein the second cell is a fibroblast, and wherein the first cell is selected from the group consisting of an astrocyte, a cardiomyocyte, and a stem cell.
21. The method of claim 1 , wherein the second cell is a neuron, and wherein the first cell is selected from the group consisting of an astrocyte and a cardiomyocyte.
22. The method of claim 1 , wherein the second cell is an astrocyte and wherein the first cell is a cardiomyocyte.
23. The method of claim 1 , wherein the second cell is cultured for at least 5 days after transfection of the mRNA transcriptome.