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Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9
JoVE Journal
Genetics
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JoVE Journal Genetics
Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

DOI:
10.3791/58130-v

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14:48 min

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August 25, 2018

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, , , , ,
1Allen Institute for Cell Science

Chapters

  • 00:00Title
  • 02:29Ribonucleoprotein (RNP) Transfection for CRISPR/Cas9 Mediated Knock-in in hiPSCs
  • 05:15FACS-enrichment of putatively edited hiPSCs
  • 07:41Generating putatively edited clonal hiPSC lines
  • 10:36Cryopreservation of clonal cell lines in 96-well plate format
  • 12:21Results
  • 13:10Conclusion

Summary

Automatic Translation

Automatic Translation

Described here is a protocol for tagging endogenously expressed proteins with fluorescent tags in human induced pluripotent stem cells using CRISPR/Cas9. Putatively edited cells are enriched by fluorescence activated cell sorting and clonal cell lines are generated.

Tags

Keywords: CRISPR/Cas9Endogenous Protein TaggingHuman Induced Pluripotent Stem CellsRibonucleoproteinFluorescent TagFACSClonal Cell LinesGene EditingLive Cell ImagingCellular StructuresCellular Processes

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