Methods for modulating embryonic stem cell differentiation

Abstract

Described herein is Zscan4, a gene exhibiting 2-cell embryonic stage and embryonic stem cell specific expression. Identification of nine Zscan4 co-expressed genes is also described Inhibition of Zscan4 expression inhibits the 2-cell to 4-cell embryonic transition and prevents blastocyst implantation, expansion and outgrowth. Provided herein are methods of inhibiting differentiation of a stem cell, promoting blastocyst outgrowth of embryonic stem cells and identifying a subpopulation of stem cells expressing Zscan4. Further described is the identification of Trim43 as a gene exhibiting morula-specific expression. Also provided are isolated expression vectors comprising a Zscan4 promoter, or a Trim43 promoter operably linked to a heterologous polypeptide and uses thereof. Further provided are transgenic animals comprising transgenes encoding marker proteins operably linked to Zscan4 and Trim43 promoters.

1 . An in vitro method of increasing the expression of a Zscan4 protein in an embryonic stem cell comprising introducing a nucleic acid encoding the Zscan4 protein into the embryonic stem cell. 
     
     
         2 . The method of  claim 1 , wherein the nucleic acid is operably linked to a promoter. 
     
     
         3 . The method of  claim 2 , wherein the nucleic acid encoding the Zscan4 protein comprises SEQ ID NO: 60. 
     
     
         4 . The method of  claim 2 , wherein the nucleic acid encoding the Zscan4 protein is at least 95% identical to SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 25, or SEQ ID NO: 29. 
     
     
         5 . The method of  claim 1 , further comprising a vector comprising the nucleic acid encoding the Zscan4 protein.