Impact of chromatin context on Cas9-induced DNA double-strand break repair pathway balance

被引：118

作者：

Schep, Ruben ^{[1
,2
]}

Brinkman, Eva K. ^{[1
,2
,7
]}

Leemans, Christ ^{[1
,2
]}

Vergara, Xabier ^{[1
,2
,3
]}

van der Weide, Robin H. ^{[1
,2
]}

Morris, Ben ^{[4
,5
]}

van Schaik, Tom ^{[1
,2
]}

Manzo, Stefano G. ^{[1
,2
]}

Peric-Hupkes, Daniel ^{[1
,2
]}

van den Berg, Jeroen ^{[1
,3
]}

Beijersbergen, Roderick L. ^{[4
,5
]}

Medema, Rene H. ^{[1
,3
]}

van Steensel, Bas ^{[1
,2
,6
]}

机构：

[1] Netherlands Canc Inst, Oncode Inst, NL-1066 CX Amsterdam, Netherlands

[2] Netherlands Canc Inst, Div Gene Regulat, NL-1066 CX Amsterdam, Netherlands

[3] Netherlands Canc Inst, Div Cell Biol, NL-1066 CX Amsterdam, Netherlands

[4] Netherlands Canc Inst, Div Mol Carcinogenesis, NL-1066 CX Amsterdam, Netherlands

[5] Netherlands Canc Inst, Robot Screening Ctr, NL-1066 CX Amsterdam, Netherlands

[6] Erasmus MC, Dept Cell Biol, NL-3015 CN Rotterdam, Netherlands

[7] Karolinska Inst, Dept Microbiol Tumor & Cell Biol, SciLifeLab, Box 1031, S-17121 Stockholm, Sweden

来源：

MOLECULAR CELL | 2021年 / 81卷 / 10期

基金：

欧洲研究理事会;

关键词：

HOMOLOGOUS RECOMBINATION REPAIR; END RESECTION; GENOME; CRISPR-CAS9; HETEROCHROMATIN; DAMAGE; HP1; RECRUITMENT; ALIGNMENT; DYNAMICS;

D O I：

10.1016/j.molcel.2021.03.032

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：

DNA double-strand break (DSB) repair is mediated by multiple pathways. It is thought that the local chromatin context affects the pathway choice, but the underlying principles are poorly understood. Using a multiplexed reporter assay in combination with Cas9 cutting, we systematically measure the relative activities of three DSB repair pathways as a function of chromatin context in >1,000 genomic locations. This reveals that non-homologous end-joining (NHEJ) is broadly biased toward euchromatin, while the contribution of microhomology-mediated end-joining (MMEJ) is higher in specific heterochromatin contexts. In H3K27me3-marked heterochromatin, inhibition of the H3K27 methyltransferase EZH2 reverts the balance toward NHEJ. Single-stranded template repair (SSTR), often used for precise CRISPR editing, competes with MMEJ and is moderately linked to chromatin context. These results provide insight into the impact of chromatin on DSB repair pathway balance and guidance for the design of Cas9-mediated genome editing experiments.

引用

页码：2216 / +

页数：25

共 102 条

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