Revealing hidden complexities of genomic rearrangements generated with Cas9

被引：43

作者：

Boroviak, Katharina ^{[1
]}

Fu, Beiyuan ^{[1
]}

Yang, Fengtang ^{[1
]}

Doe, Brendan ^{[1
]}

Bradley, Allan ^{[1
]}

机构：

[1] Wellcome Trust Sanger Inst, Wellcome Genome Campus, Cambridge CB10 1SA, England

来源：

SCIENTIFIC REPORTS | 2017年 / 7卷

基金：

英国惠康基金;

关键词：

IMMUNE-SYSTEM; MOUSE MODEL; MICE; CRISPR/CAS; EFFICIENT;

D O I：

10.1038/s41598-017-12740-6

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

摘要：

Modelling human diseases caused by large genomic rearrangements has become more accessible since the utilization of CRISPR/Cas9 in mammalian systems. In a previous study, we showed that genomic rearrangements of up to one million base pairs can be generated by direct injection of CRISPR/Cas9 reagents into mouse zygotes. Although these rearrangements are ascertained by junction PCR, we describe here a variety of anticipated structural changes often involving reintegration of the region demarcated by the gRNAs in the vicinity of the edited locus. We illustrate here some of this diversity detected by high-resolution fibre-FISH and conclude that extensive molecular analysis is required to fully understand the structure of engineered chromosomes generated by Cas9.

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