Generation of a knock-in MAP2-tdTomato reporter human embryonic stem cell line with inducible expression of NEUROG2/1 (NYGCe001-A)

Stem Cell Res. 2019 Dec:41:101643. doi: 10.1016/j.scr.2019.101643. Epub 2019 Nov 1.

Abstract

Overexpression of NEUROG2 and NEUROG1 (NEUROG2/1) in human embryonic stem cells (hESCs) rapidly produces functional networks of excitatory and inhibitory neurons. To facilitate the use of this efficient inducible human neuron model in neuroscience research, we generated hESCs with doxycycline-inducible NEUROG2/1 via lentivirus and a tdTomato fluorescent reporter knock-in at the MAP2 locus using the CRISPR nuclease Cas9. Upon doxycycline-driven induction of NEUROG2/1, these hESCs differentiate within days into cells that are uniformly MAP2 immunoreactive and tdTomato fluorescent.

Publication types

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

MeSH terms

  • Base Sequence
  • Basic Helix-Loop-Helix Transcription Factors / metabolism*
  • Cell Culture Techniques / methods*
  • Cell Line / cytology*
  • Female
  • Gene Knock-In Techniques*
  • Genes, Reporter*
  • Human Embryonic Stem Cells / metabolism*
  • Humans
  • Microtubule-Associated Proteins / metabolism*
  • Nerve Tissue Proteins / metabolism*
  • Reproducibility of Results

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • MAP2 protein, human
  • Microtubule-Associated Proteins
  • NEUROG1 protein, human
  • NEUROG2 protein, human
  • Nerve Tissue Proteins