title: Inhibition of master transcription factors in pluripotent cells induces early stage differentiation.
abstract: The potential for pluripotent cells to differentiate into diverse specialized cell types has given much hope to the field of regenerative medicine. Nevertheless, the low efficiency of cell commitment has been a major bottleneck in this field. Here we provide a strategy to enhance the efficiency of early differentiation of pluripotent cells. We hypothesized that the initial phase of differentiation can be enhanced if the transcriptional activity of master regulators of stemness is suppressed, blocking the formation of functional transcriptomes. However, an obstacle is the lack of an efficient strategy to block protein-protein interactions. In this work, we take advantage of the biochemical property of seventeen kilodalton protein (Skp), a bacterial molecular chaperone that binds directly to sex determining region Y-box 2 (Sox2). The small angle X-ray scattering analyses provided a low resolution model of the complex and suggested that the transactivation domain of Sox2 is probably wrapped in a cleft on Skp trimer. Upon the transduction of Skp into pluripotent cells, the transcriptional activity of Sox2 was inhibited and the expression of Sox2 and octamer-binding transcription factor 4 was reduced, which resulted in the expression of early differentiation markers and appearance of early neuronal and cardiac progenitors. These results suggest that the initial stage of differentiation can be accelerated by inhibiting master transcription factors of stemness. This strategy can possibly be applied to increase the efficiency of stem cell differentiation into various cell types and also provides a clue to understanding the mechanism of early differentiation.
authors.affiliation: Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon 440-746, Korea.
authors.forename: Debojyoti
authors.initials: D
authors.lastname: De
authors.forename: Myong-Ho
authors.initials: MH
authors.lastname: Jeong
authors.forename: Young-Eun
authors.initials: YE
authors.lastname: Leem
authors.forename: Dmitri I
authors.initials: DI
authors.lastname: Svergun
authors.forename: David E
authors.initials: DE
authors.lastname: Wemmer
authors.forename: Jong-Sun
authors.initials: JS
authors.lastname: Kang
authors.forename: Kyeong Kyu
authors.initials: KK
authors.lastname: Kim
authors.forename: Sung-Hou
authors.initials: SH
authors.lastname: Kim
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D029968:Escherichia coli Proteins / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D008958:Models, Molecular; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D053838:Scattering, Small Angle; D012996:Solutions; D014157:Transcription Factors / Q000378:metabolism*; D014161:Transduction, Genetic; D014961:X-Ray Diffraction; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D055748:SOXB1 Transcription Factors; D012996:Solutions; D014157:Transcription Factors; D054322:tat Gene Products, Human Immunodeficiency Virus