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CRISPR/Cas9 诱导人多潜能干细胞内源蛋白标记的应用
JoVE Journal
Genética
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JoVE Journal Genética
Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9
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CRISPR/Cas9 诱导人多潜能干细胞内源蛋白标记的应用

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

Capítulos

  • 00:00Título
  • 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
  • 12:21Resultados
  • 13:10Conclusion

Summary

Tadução automática

Tadução automática

这里描述的是一个协议, 用于标记内在表达的蛋白与荧光标记在人类诱导多潜能干细胞使用 CRISPR/Cas9。认定编辑细胞通过荧光活化细胞分选, 生成克隆细胞系。

Tags

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

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