Journal
/
Genetics
/
在人类多能干细胞中使用CRISPR-CAS9生成定义的基因组修饰
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
Please note that all translations are automatically generated. Click here for the English version.

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

•

09:04 min

•

September 25, 2019

•

, , ,
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

Automatic Translation

Automatic Translation

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

用荧光PCR毛细管电泳技术在基因型CRISPR / Cas9介导的敲除突变体的高通量形式

选择依赖和独立生成CRISPR / Cas9介导的哺乳动物细胞基因敲除

CRISPR/Cas9-based 跳跃的原始生殖细胞移植在爪

DNA 修饰的免疫: 共焦图像的计算分析

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

CRISPR/Cas9 对小鼠和人造血祖细胞的高效基因破坏

利用 CRISPR/Cas9 Ribonucleoproteins 快速简便地生成线虫基因组点突变体的管道

CRISPR/Cas9 基因编辑制作人类疟疾寄生虫的条件突变体

CRISPR/Cas9 诱导人多潜能干细胞内源蛋白标记的应用

慢病毒载体平台, 用于将表观基因编辑工具高效地传递到人类诱导的多能干细胞衍生疾病模型中

Read Article