Methods for generating cardiomyocytes
The present disclosure provides method of generating cardiomyocytes from post-natal fibroblasts. The present disclosure further provides cells and compositions for use in generating cardiomyocytes.
1. A method of generating an induced cardiomyocyte, the method comprising: introducing into a human post-natal fibroblast reprogramming factors comprising Gata4, Mef2c, Tbx5, Mesp1, and Esrrg, wherein said introducing results in direct reprogramming of the post-natal fibroblast into a cardiomyocyte, thereby generating an induced cardiomyocyte.
2. The method of claim 1 , wherein said introducing comprises genetically modifying the post-natal fibroblast with one or more nucleic acids comprising nucleotide sequences encoding the reprogramming factor polypeptides.
3. The method of claim 2 , wherein said nucleic acid is a recombinant vector.
4. The method of claim 2 , wherein the nucleotide sequences are operably linked to a transcription regulatory element.
5. The method of claim 1 , wherein said reprogramming factors are exogenous, and where said introducing comprises introducing the exogenous reprogramming factor polypeptides into the post-natal fibroblast.
6. The method of claim 1 , wherein said introducing is carried out in vitro.
7. The method of claim 1 , wherein said introducing is carried out in vivo.
8. The method of claim 1 , wherein the Esrrg factor comprises an amino acid sequence having at least 80% amino acid identity to SEQ ID NO:1.
9. The method of claim 1 , wherein the Gata4 factor comprises an amino acid sequence having at least 80% amino acid identity to SEQ ID NO:8.
10. The method of claim 1 , wherein the Mef2c factor comprises an amino acid sequence having at least 80% amino acid identity to SEQ ID NO:10 or SEQ ID NO:12.
11. The method of claim 1 , wherein the Tbx5 factor comprises an amino acid sequence having at least 80% amino acid identity to SEQ ID NO:14 or SEQ ID NO:16.
12. The method of claim 1 , wherein the Mesp1 factor comprises an amino acid sequence having at least 80% amino acid identity to SEQ ID NO:18.
13. The method of claim 1 , wherein the method does not comprise introducing into the human post-natal fibroblast an exogenous Sox2 polypeptide or a nucleic acid encoding the exogenous Sox2 polypeptide.