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CRISPR/Cas9-mediated Precise SNP Editing in Human iPSC Lines.


ABSTRACT: Human induced pluripotent stem cells (hiPSCs) have been extensively used in the fields of developmental biology and disease modeling. CRISPR/Cas9 gene editing in iPSC lines often has a low frequency, which hampers its application in precise allele editing of disease-associated single nucleotide polymorphisms (SNPs), especially those in the noncoding parts of the genome. Here, we present a unique workflow to engineer isogenic iPSC lines by SNP editing from heterozygous to homozygous for disease risk alleles or non-risk alleles using a transient and straightforward transfection-based protocol. This protocol enables us to simultaneously obtain pure and clonal isogenic lines of all three possible genotypes of a SNP site within about 4 to 5 weeks.

SUBMITTER: Zhang H 

PROVIDER: S-EPMC8260261 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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CRISPR/Cas9-mediated Precise SNP Editing in Human iPSC Lines.

Zhang Hanwen H   Zhang Siwei S  

Bio-protocol 20210620 12


Human induced pluripotent stem cells (hiPSCs) have been extensively used in the fields of developmental biology and disease modeling. CRISPR/Cas9 gene editing in iPSC lines often has a low frequency, which hampers its application in precise allele editing of disease-associated single nucleotide polymorphisms (SNPs), especially those in the noncoding parts of the genome. Here, we present a unique workflow to engineer isogenic iPSC lines by SNP editing from heterozygous to homozygous for disease r  ...[more]

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