Knockdown of Oct-4 or Sox-2 attenuates neurogenesis of mouse embryonic stem cells.
We employed a stromal-derived inducing activity (SDIA) model of neurogenesis to investigate the effects of targeted knockdown of Oct-4 and Sox-2 by short interfering RNAs (siRNAs) in mouse embryonic stem (mES) cells. Quantitative real-time PCR showed 40-90% knockdown of specific transcripts with cognate Oct-4 or Sox-2 siRNA transfection compared to FAM-labeled negative control (FAM) siRNA or mock transfection and was confirmed at the protein level by western blot analyses. Upon differentiation using PA6 SDIA co-cultures, neurogenesis is significantly diminished in Oct-4 or Sox-2-targeted mES cells. It was observed that 45 +/- 12%, 65 +/- 13%, and 90 +/- 8% of the colonies were stained with neuron-specific beta-tubulin III in Oct-4, Sox-2, and FAM siRNA transfected mES cells, respectively, with similar results observed using neural inducing factors collected from the surface of PA6. Together, our results extend observations for a role of Oct-4 in SDIA and implicate a similar role for Sox-2.
Chen
Stephen
S
Stem Cell Group, Bioprocessing Technology Institute, Centros, Singapore.
Choo
Andre B H
AB
Nai-Dy
Wang
W
Heng-Phon
Too
T
Oh
Steve K W
SK
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D018920:Coculture Techniques; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation; D024002:HMGB Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
D015415:Biomarkers; D004268:DNA-Binding Proteins; D024002:HMGB Proteins; D050814:Octamer Transcription Factor-3; D034741:RNA, Small Interfering; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D014157:Transcription Factors