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使用 CRISPR 介质基础编辑器对 BRCA1 变体进行功能评估

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Cancer Research

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JoVE Journal Cancer Research

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Please note that all translations are automatically generated. Click here for the English version.

使用 CRISPR 介质基础编辑器对 BRCA1 变体进行功能评估

DOI:

10.3791/61557-v

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09:22 min

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February 28, 2021

•

Ji-Eun See², Ha Rim Shin², Gayoung Jang², Jiyeon Kweon², Yongsub Kim²

¹Department of Biomedical Sciences, Asan Medical Institute of Convergence Science and Technology, Asan Medical Center,University of Ulsan College of Medicine, ²Stem Cell Immunomodulation Research Center,University of Ulsan College of Medicine

Chapters

00:05Introduction
01:03Design and Construction of BRCA1 Targeting gRNAs
03:17Creation of BRCA1 Variants using CRISPR-mediated Base Editing Tools
04:15Sample Preparation for Illumina Next-Generation Sequencing (NGS)
07:01Results: Functional Study of BRCA1 using CRISPR-Cas9 Systems
08:34Conclusion

Summary

Automatic Translation

BRCA1突变患者患癌症的风险较高，这值得准确评估BRCA1变异的功能。在此，我们描述了使用CRISPR介质细胞素基础编辑器对BRCA1变种进行功能评估的协议，该基编辑器支持活细胞中目标C：G到T：A的转换。

Tags

BRCA1 CRISPR Base Editors Functional Assessment Point Mutations Cancer Drug Targets Genome Sequence Guide RNA Oligonucleotides Plasmid DNA Transformation Sanger Sequencing HAP1-BE3 Cells Transfection

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