Induced pluripotent stem cells

Abstract

Described herein is a major breakthrough in nuclear reprogramming and induced pluripotent stem cell (iPSC) technology. Fusion of the powerful transcription activation domain (TAD) of MyoD to the Oct4 protein makes iPSCs generation faster, more efficient, purer, safer and feeder-free. Also, disclosed herein is the first report of the use of a TAD fused to a transcription factor as a method for making iPSCs. By combining transcription factors and TADs, this approach to nuclear reprogramming can have a range of applications from inducing pluriopotency to inducing transdifferentiation without transitioning through iPSCs.

1 . A composition comprising a nucleic acid encoding a fusion protein comprising a transactivation domain from MyoD or VP16 fused to the N-terminus or C-terminus of Oct4 operably linked to a promoter. 
     
     
         2 . The composition of  claim 1 , further comprising at least one nucleic acid encoding Sox2, Klf4, and optionally c-Myc. 
     
     
         3 . The composition of  claim 1 , wherein the transactivation domain is obtained from MyoD. 
     
     
         4 . The composition of  claim 3 , wherein the transactivation domain of MyoD comprises an N-terminus region of MyoD. 
     
     
         5 . The composition of  claim 4 , wherein the transactivation domain comprising amino acids 1-62 of MyoD or is at least 95% identical thereto. 
     
     
         6 .- 19 . (canceled) 
     
     
         20 . The composition of  claim 1 , wherein said composition is contained within a cell. 
     
     
         21 . The composition of  claim 20 , wherein said cell is a mammalian cell. 
     
     
         22 . The composition of  claim 21 , wherein said mammalian cell is human. 
     
     
         23 . The composition of  claim 2 , wherein said composition is contained within a cell. 
     
     
         24 . The composition of  claim 23 , wherein said cell is a mammalian cell. 
     
     
         25 . The composition of  claim 24 , wherein said mammalian cell is human. 
     
     
         26 . The composition of  claim 2 , wherein the nucleic acid is polycistronic. 
     
     
         27 . The composition of  claim 2 , wherein the proteins encoded by said at least one nucleic acid are human. 
     
     
         28 . The composition of  claim 2 , further comprising a cytokine. 
     
     
         29 . The composition of  claim 28 , wherein the cytokine is basic fibroblast growth factor and/or stem cell factor.