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

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

DOI:
10.3791/61152-v

•

09:03 min

•

May 10, 2020

•

, ,
1MRC再生医学センター,University of Edinburgh

章

  • 00:05Introduction
  • 00:40Gene Editing of Human Pluripotent Stem Cells
  • 06:57Results: Genetic Editing in Human Pluripotent Stem Cells
  • 08:06Conclusion

概要

自動翻訳

自動翻訳

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).

タグ

Human Pluripotent Stem CellsGenome EditingCRISPR-Cas9Seamless Genome EditingTargeted Gene InsertionLaminin 521MTeSR1 MediumCell DissociationNucleofectionSingle Guide RNATargeting Vector

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