A high efficiency precision genome editing method with CRISPR in iPSCs
Avinash Singh, G. Dalton Smedley, J. Rose, Kristina Fredriksen, Ying Zhang, Ling Li, Shauna H. Yuan
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.
Topics & Concepts
CRISPRInduced pluripotent stem cellGenome editingPalindromeHomologous recombinationComputational biologyBiologyPoint mutationHuman Induced Pluripotent Stem CellsGenomeGeneticsGeneMutationEmbryonic stem cellCRISPR and Genetic EngineeringPluripotent Stem Cells ResearchBiomedical Ethics and Regulation