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	A novel technique based on in vitro oocyte injection to improve CRISPR/Cas9 gene editing in zebrafish
	Xie, Shao-Lin1,2; Bian, Wan-Ping2; Wang, Chao1,2; Junaid, Muhammad2,3; Zou, Ji-Xing1; Pei, De-Sheng2
	2016-09-29
摘要	Contemporary improvements in the type II clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system offer a convenient way for genome editing in zebrafish. However, the low efficiencies of genome editing and germline transmission require a time-intensive and laborious screening work. Here, we reported a method based on in vitro oocyte storage by injecting oocytes in advance and incubating them in oocyte storage medium to significantly improve the efficiencies of genome editing and germline transmission by in vitro fertilization (IVF) in zebrafish. Compared to conventional methods, the prior micro-injection of zebrafish oocytes improved the efficiency of genome editing, especially for the sgRNAs with low targeting efficiency. Due to high throughputs, simplicity and flexible design, this novel strategy will provide an efficient alternative to increase the speed of generating heritable mutants in zebrafish by using CRISPR/Cas9 system.
DOI	10.1038/srep34555
URL	查看原文
发表期刊	SCIENTIFIC REPORTS
ISSN	2045-2322
卷号	6页码:10
通讯作者	Zou, JX (reprint author), South China Agr Univ, Coll Marine Sci, Guangzhou 510642, Guangdong, Peoples R China. ; Pei, DS (reprint author), Chinese Acad Sci, Chongqing Inst Green & Intelligent Technol, Chongqing 400714, Peoples R China.
收录类别	SCI
WOS记录号	WOS:000384193200001
语种	英语