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在人类多能干细胞中使用CRISPR-CAS9生成定义的基因组修饰
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Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
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在人类多能干细胞中使用CRISPR-CAS9生成定义的基因组修饰

DOI:
10.3791/60085-v

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09:04 min

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September 25, 2019

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, , ,
1Center for Cellular and Molecular Therapeutics,The Children’s Hospital of Philadelphia, 2Department of Pathology and Laboratory Medicine,The Children’s Hospital of Philadelphia, 3Department of Pathology and Laboratory Medicine,University of Pennsylvania

Chapters

  • 00:04Title
  • 01:19Test gRNA (GuidE RNA) Cutting Efficiency
  • 03:50Clone Screening
  • 04:51Transfection Setup
  • 06:07Checking for Mutations in Single Colonies
  • 07:11Results: Cutting Efficiency, and Generation and Screening of Mutations
  • 08:10Conclusion

Summary

자동 번역

자동 번역

该协议提供了一种方法，促进在人类多能干细胞中使用CRISPR-CAS9生成定义的异质核苷酸或同源核苷酸变化。

Tags

CRISPR-Cas9Human Pluripotent Stem CellsIsogenic LinesHeterozygous MutationsGene FunctionDisease ModelingColony PickingFluorescence-activated Cell SortingMEFsROCK Inhibitor

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