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Published November 2004 | public
Journal Article

Directing pluripotent cell differentiation using "diced RNA" in transient transfection

Abstract

Embryonic stem (ES) and embryonic carcinoma (EC) cells are pluripotent and have the capacity to differentiate into many cell types. The ability to direct their differentiation should have considerable practical applications. Here, we first report the use of diced short interfering RNAi against Oct4 in a transient approach, to direct differentiation of ES towards the trophectoderm lineage. We then apply this approach to downregulate Smad4 in mouse P19 EC cells. We have found that this leads to an increase in the levels of Pax6 (a neuroectoderm marker), reduction in the levels of Brachyury (a mesoderm marker), and a 3‐fold increase in the number of βIII tubulin‐positive colonies when these cells were allowed to differentiate. This indicates a redirection of cell fate towards the neuroectoderm lineage. Thus, transient RNAi could provide a valuable tool to direct pluripotent cells along specific pathways of differentiation while circumventing permanent genetic changes.

Additional Information

© 2004 Wiley‐Liss. Received 19 April 2004; Accepted 15 July 2004. Contract grant sponsor: Cancer Research UK (CRUK), Contract grant number: C3/A2943. We thank Neil Clarke, Chris Ring, and David Glover for valuable discussions and critical review of this paper. MZG is a Wellcome Trust Senior Research Fellow.

Additional details

Created:
August 22, 2023
Modified:
October 20, 2023