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Genetics

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JoVE Journal Genetics
Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing
 

Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing

Article DOI: 10.3791/61152-v • 09:03 min • May 10th, 2020
May 10th, 2020 •
1, 1, 1
1MRC Centre for Regenerative Medicine, University of Edinburgh

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Here, we describe a detailed method for seamless gene editing in human pluripotent stem cells using a piggyBac-based donor plasmid and the Cas9 nickase mutant. Two point mutations were introduced into exon 8 of the hepatocyte nuclear factor 4 alpha (HNF4α) locus in human embryonic stem cells (hESCs).

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Point Mutations Human Pluripotent Stem Cells Genome Editing Cas-9 Targeting Vector Seamless Genomic Editing Genotyping Screening Laminin 521 MTeSR1 Medium Transfection ROCK Inhibitor Cell Dissociation Reagent
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