Generation of induced neuronal cells by the single reprogramming factor ASCL1

Stem Cell Reports. 2014 Aug 12;3(2):282-96. doi: 10.1016/j.stemcr.2014.05.020. Epub 2014 Jul 4.

Abstract

Direct conversion of nonneural cells to functional neurons holds great promise for neurological disease modeling and regenerative medicine. We previously reported rapid reprogramming of mouse embryonic fibroblasts (MEFs) into mature induced neuronal (iN) cells by forced expression of three transcription factors: ASCL1, MYT1L, and BRN2. Here, we show that ASCL1 alone is sufficient to generate functional iN cells from mouse and human fibroblasts and embryonic stem cells, indicating that ASCL1 is the key driver of iN cell reprogramming in different cell contexts and that the role of MYT1L and BRN2 is primarily to enhance the neuronal maturation process. ASCL1-induced single-factor neurons (1F-iN) expressed mature neuronal markers, exhibited typical passive and active intrinsic membrane properties, and formed functional pre- and postsynaptic structures. Surprisingly, ASCL1-induced iN cells were predominantly excitatory, demonstrating that ASCL1 is permissive but alone not deterministic for the inhibitory neuronal lineage.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials
  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism*
  • Cell Line
  • Cellular Reprogramming*
  • Embryonic Stem Cells / cytology
  • Fibroblasts / cytology
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Neural Stem Cells / cytology*
  • POU Domain Factors / genetics
  • POU Domain Factors / metabolism
  • Patch-Clamp Techniques
  • Potassium Channels / metabolism
  • Sodium Channels / metabolism
  • Synapses / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Ascl1 protein, mouse
  • Basic Helix-Loop-Helix Transcription Factors
  • Myt1l protein, mouse
  • Nerve Tissue Proteins
  • POU Domain Factors
  • Potassium Channels
  • Sodium Channels
  • Transcription Factors
  • Pou3f2 protein, mouse