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

作     者：Jia Li Hong-Yu Li Shan-Ye Gu Hua-Xing Zi Lai Jiang Jiu-Lin Du Jia Li;Hong-Yu Li;Shan-Ye Gu;Hua-Xing Zi;Lai Jiang;Jiu-Lin Du

作者机构：Institute of NeuroscienceState Key Laboratory of NeuroscienceCAS Center for Excellence in Brain Science and Intelligence TechnologyShanghai Research Center for Brain Science and Brain-Inspired IntelligenceChinese Academy of SciencesShanghai 200031China University of Chinese Academy of SciencesBeijing 100049China Department of Anesthesiology and Surgical Intensive Care UnitXinhua HospitalShanghai Jiao Tong University School of MedicineShanghai 200092China School of Life Science and TechnologyShanghai Tech UniversityShanghai 200031China 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：2020年第63卷第1期

页      面：59-67页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：supported by the Young Scientists Fund of the National Natural Science Foundation of China(31500849) Shanghai Municipal Science and Technology Major Project(18JC1410100,2018SHZDZX05) the Key Research Program of Frontier Sciences(QYZDY-SSW-SMC028) the Strategic Priority Research Program(XDB32010200)of Chinese Academy of Sciences the International Partnership Program,Bureau of International Co-operation of Chinese Academy of Sciences(153D31KYSB20170059) China Wan-Ren Program,and Shanghai Leading Scientist Program 

主　　题：NHEJ non-HR knockin conditional knockout visible switch zCKOIS zebrafish 

摘      要：The zebrafish has become a popular vertebrate animal model in biomedical ***,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 *** this strategy,we generated hey2^zCKOIS which served as a hey2 KI reporter with EGFP *** Cre induction in targeted cells,the hey2^zCKOIS was switched to a non-functional CKO allele hey2^zCKOIS-inv associated with Tag RFP expression,enabling visualization of the CKO ***,simplification of the design,and the visibility and combination of both CKO and KI alleles make our z CKOIS strategy an applicable CKO approach for zebrafish.