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Journal / Genetics / Introducing Point Mutations into Human…
JoVE Journal
Genetics

This Contenuto is Open Access.

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).

Tags

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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