Generation of two H1 hESC sublines carrying a heterozygous and homozygous knock-out of RB1

Stem Cell Res. 2017 Dec:25:270-273. doi: 10.1016/j.scr.2017.07.005. Epub 2017 Jul 11.

Abstract

Retinoblastoma is a childhood cancer of the retina caused by biallelic inactivation of the tumor suppressor gene RB1. In heritable retinoblastoma, one allele is inherited in mutant form via one of the parental germ cells. To study molecular mechanisms in retinoblastoma, two sublines of H1 hESCs were generated, carrying a knock-out allele of RB1 in the heterozygous or homozygous state. Exon 3 of RB1 was targeted and modified by nucleotide deletions using the CRISPR/Cas9 nuclease system. Based on a nearby single nucleotide polymorphism, the modification could be assigned to one allele.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cells, Cultured
  • Gene Knockout Techniques
  • Heterozygote
  • Homozygote
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / metabolism
  • Humans
  • Mutation
  • Retinoblastoma / embryology
  • Retinoblastoma / genetics*
  • Retinoblastoma / metabolism
  • Retinoblastoma Protein / genetics*
  • Retinoblastoma Protein / metabolism

Substances

  • Retinoblastoma Protein