A CRISPR-based base-editing screen for the functional assessment of BRCA1 variants

被引：65

作者：

Kweon, Jiyeon ^{[1
,2
]}

Jang, An-Hee ^{[1
,2
]}

Shin, Ha Rim ^{[1
,2
]}

See, Ji-Eun ^{[1
,2
]}

Lee, Woochang ^{[3
]}

Lee, Jong Won ^{[4
]}

Chang, Suhwan ^{[1
]}

Kim, Kyunggon ^{[5
,6
]}

Kim, Yongsub ^{[1
,2
]}

机构：

[1] Univ Ulsan, Coll Med, Asan Med Inst Convergence Sci & Technol, Dept Biomed Sci,Asan Med Ctr, Seoul, South Korea

[2] Univ Ulsan, Coll Med, Stem Cell Immunomodulat Res Ctr, Seoul, South Korea

[3] Univ Ulsan, Coll Med, Asan Med Ctr, Dept Lab Med, Seoul, South Korea

[4] Univ Ulsan, Coll Med, Asan Med Ctr, Div Breast Surg,Dept Surg, Seoul, South Korea

[5] Univ Ulsan, Coll Med, Asan Med Ctr, Dept Convergence Med, Seoul, South Korea

[6] Asan Inst Life Sci, Clin Prote Core Lab, Seoul, South Korea

来源：

ONCOGENE | 2020年 / 39卷 / 01期

基金：

新加坡国家研究基金会;

关键词：

GENOMIC DNA; SITES;

D O I：

10.1038/s41388-019-0968-2

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Genetic mutations in BRCA1, which is crucial for the process of DNA repair and maintenance of genomic integrity, are known to increase markedly the risk of breast and ovarian cancers. Clinical genetic testing has been used to identify new BRCA1 variants; however, functional assessment and determination of their pathogenicity still poses challenges for clinical management. Here, we describe that CRISPR-mediated cytosine base editor, known as BE3, can be used for the functional analysis of BRCA1 variants. We performed CRISPR-mediated base-editing screening using 745 gRNAs targeting all exons in BRCA1 to identify loss-of-function variants and identified variants whose function has heretofore remained unknown, such as c.-97C>T, c.154C>T, c.3847C>T, c.5056C>T, and c.4986+5G>A. Our results show that CRISPR-mediated base editor is a powerful tool for the reclassification of variants of uncertain significance (VUSs) in BRCA1.

引用

页码：30 / 35

页数：6

共 26 条

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