One-step generation of zebrafish carrying a conditional knockout-knockin visible switch via CRISPR/Cas9-mediated intron targeting

doi:10.1007/s11427-019-1607-9

	One-step generation of zebrafish carrying a conditional knockout-knockin visible switch via CRISPR/Cas9-mediated intron targeting
	Li, Jia 1; Li, Hong-Yu 1,2; Gu, Shan-Ye 3,4; Zi, Hua-Xing 1,2; Jiang, Lai 3,4; Du, Jiu-Lin1,2,5
	2019-12-20
发表期刊	SCIENCE CHINA-LIFE SCIENCES
ISSN	1674-7305
EISSN	1869-1889
发表状态	已发表
DOI	10.1007/s11427-019-1607-9
摘要	The zebrafish has become a popular vertebrate animal model in biomedical research. However, it is still challenging to make conditional gene knockout (CKO) models in zebrafish due to the low efficiency of homologous recombination (HR). Here we report an efficient non-HR-based method for generating zebrafish carrying a CKO and knockin (KI) switch (zCKOIS) coupled with dual-color fluorescent reporters. Using this strategy, we generated hey2(zKOIS) which served as a hey2 KI reporter with EGFP expression. Upon Cre induction in targeted cells, the hey2(zCKOIS) was switched to a non-functional CKO allele hey2(zCKOIS-inv)associated with TagRFP expression, enabling visualization of the CKO alleles. Thus, simplification of the design, and the visibility and combination of both CKO and KI alleles make our zCKOIS strategy an applicable CKO approach for zebrafish.
关键词	NHEJ non-HR knockin conditional knockout visible switch zCKOIS zebrafish
URL	查看原文
收录类别	SCI ; SCIE
语种	英语
资助项目	International Partnership Program, Bureau of International Co-operation of Chinese Academy of Sciences[153D31KYSB20170059]
WOS研究方向	Life Sciences & Biomedicine - Other Topics
WOS类目	Biology
WOS记录号	WOS:000504160900003
出版者	SCIENCE PRESS
WOS关键词	GRIDLOCK ; GENOME ; CAS9
原始文献类型	Article ; Early Access
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/104983
专题	生命科学与技术学院_特聘教授组_杜久林组
通讯作者	Li, Jia; Du, Jiu-Lin
作者单位	1.Chinese Acad Sci, Shanghai Res Ctr Brain Sci & Brain Inspired Intel, CAS Ctr Excellence Brain Sci & Intelligence Techn, Inst Neurosci,State Key Lab Neurosci, Shanghai 200031, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Shanghai Jiao Tong Univ, Sch Med, Xinhua Hosp, Dept Anesthesiol, Shanghai 200092, Peoples R China 4.Shanghai Jiao Tong Univ, Sch Med, Xinhua Hosp, Surg Intens Care Unit, Shanghai 200092, Peoples R China 5.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 200031, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Li, Jia,Li, Hong-Yu,Gu, Shan-Ye,et al. One-step generation of zebrafish carrying a conditional knockout-knockin visible switch via CRISPR/Cas9-mediated intron targeting[J]. SCIENCE CHINA-LIFE SCIENCES,2019.
APA	Li, Jia,Li, Hong-Yu,Gu, Shan-Ye,Zi, Hua-Xing,Jiang, Lai,&Du, Jiu-Lin.(2019).One-step generation of zebrafish carrying a conditional knockout-knockin visible switch via CRISPR/Cas9-mediated intron targeting.SCIENCE CHINA-LIFE SCIENCES.
MLA	Li, Jia,et al."One-step generation of zebrafish carrying a conditional knockout-knockin visible switch via CRISPR/Cas9-mediated intron targeting".SCIENCE CHINA-LIFE SCIENCES (2019).