Generation of a FMR1 homozygous knockout human embryonic stem cell line (WAe009-A-16) by CRISPR/Cas9 editing

Stem Cell Res. 2019 Aug:39:101494. doi: 10.1016/j.scr.2019.101494. Epub 2019 Jun 28.

Abstract

Mutations in FMR1 gene is the cause of Fragile X Syndrome (FXS) leading inherited cause of intellectual disability and autism spectrum disorders. FMR1 gene encodes Fragile X Mental Retardation Protein (FMRP) which is a RNA binding protein and play important role in synaptic plasticity and translational regulation in neurons. We have generated a homozygous FMR1 knockout (FMR1-KO) hESC line using CRISPR/Cas9 based genome editing. It created a homozygous 280 nucleotide deletion at exon1, removing the start codon. This FMR1-KO cell line maintains stem cell like morphology, pluripotency, normal karyotype and ability to in-vitro differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • CRISPR-Cas Systems / genetics*
  • Cell Differentiation / genetics
  • Cell Differentiation / physiology
  • Cell Line
  • Embryoid Bodies / cytology
  • Embryoid Bodies / metabolism
  • Exons / genetics
  • Fragile X Mental Retardation Protein / genetics*
  • Genotype
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / metabolism*
  • Humans
  • Immunohistochemistry
  • Karyotype
  • RNA-Binding Proteins / genetics

Substances

  • FMR1 protein, human
  • RNA-Binding Proteins
  • Fragile X Mental Retardation Protein