Creating a patient carried Men1 gene point mutation on wild type iPSCs locus mediated by CRISPR/Cas9 and ssODN

Stem Cell Res. 2017 Jan:18:67-69. doi: 10.1016/j.scr.2016.12.007. Epub 2016 Dec 9.

Abstract

A patient specific point mutation (c.1288G>T) of Men1 gene was introduced into wide type iPSC line with CRISPR/Cas9 and single-stranded donor oligonucleotides carrying the mutation. The mutated iPSC line has a heterozygous c.1288G>T mutation on exon-9 of Men1 that was confirmed by sequencing analysis. The karyotype of this line was normal and the pluripotency was demonstrated by its ability to differentiate into three germ layers. These artificially created Men1 mutation in wild type iPSC line will help to dissect out the molecular basis of two patients carried the same mutation from one family who were differentially represented hypoglycemia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • CRISPR-Cas Systems / genetics*
  • Cell Differentiation
  • Cell Line
  • DNA Mutational Analysis
  • Embryoid Bodies / metabolism
  • Embryoid Bodies / pathology
  • Exons
  • Gene Editing
  • Genotype
  • Heterozygote
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Karyotype
  • Male
  • Microscopy, Fluorescence
  • Multiple Endocrine Neoplasia Type 1 / genetics
  • Multiple Endocrine Neoplasia Type 1 / metabolism
  • Multiple Endocrine Neoplasia Type 1 / pathology
  • Oligodeoxyribonucleotides / genetics*
  • Oligodeoxyribonucleotides / metabolism
  • Point Mutation
  • Proto-Oncogene Proteins / genetics*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • MEN1 protein, human
  • Oligodeoxyribonucleotides
  • Proto-Oncogene Proteins
  • Transcription Factors