Generation of a SMO homozygous knockout human embryonic stem cell line WAe001-A-16 by CRISPR/Cas9 editing

Stem Cell Res. 2018 Mar:27:5-9. doi: 10.1016/j.scr.2017.12.002. Epub 2017 Dec 5.

Abstract

The human SMO protein encoded by the smoothened (SMO) gene acts as a positive mediator for Hedgehog signaling. This pathway regulates many cellular activities, developmental morphogenesis, and tumorigenesis. Using CRISPR/Cas9 to edit human embryonic stem cell line WA01 (H1), we established a SMO mutant cell line (WAe001-A-16). This cell line has a 40bp homozygous deletion in exon 2 of SMO leading to a shift in the open reading frame and early termination at amino acid position 287. WAe001-A-16 maintains a normal karyotype, parental cell morphology, pluripotency markers, and the capacity to differentiate into all three germline layers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems / genetics
  • CRISPR-Cas Systems / physiology
  • Cell Differentiation / genetics
  • Cell Differentiation / physiology
  • Cell Line
  • Cilia / genetics
  • Cilia / metabolism
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism*
  • Human Embryonic Stem Cells / metabolism*
  • Humans
  • Karyotype
  • Open Reading Frames / genetics
  • Smoothened Receptor / genetics
  • Smoothened Receptor / metabolism*

Substances

  • SMO protein, human
  • Smoothened Receptor