A high efficiency precision genome editing method with CRISPR in iPSCs

Sci Rep. 2024 Apr 30;14(1):9933. doi: 10.1038/s41598-024-60766-4.

Abstract

The use of genetic engineering to generate point mutations in induced pluripotent stem cells (iPSCs) is essential for studying a specific genetic effect in an isogenic background. We demonstrate that a combination of p53 inhibition and pro-survival small molecules achieves a homologous recombination rate higher than 90% using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) in human iPSCs. Our protocol reduces the effort and time required to create isogenic lines.

Keywords: CRISPR; Gene editing; High efficiency; Single nucleotide polymorphism; iPSC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems*
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Gene Editing* / methods
  • Homologous Recombination
  • Humans
  • Induced Pluripotent Stem Cells* / cytology
  • Induced Pluripotent Stem Cells* / metabolism
  • Tumor Suppressor Protein p53 / genetics

Substances

  • Tumor Suppressor Protein p53