GoldenFish:a rapid and efficient system to customize constructs for zebrafish transgenesis  

作　　者：Zhanmei Jiang Lirong Huang Jieqiong Zhao Yanfeng Li Jianlong Ma Rui Ni Qifen Yang Lingfei Luo Yun Yang Jingying Chen 

机构地区：[1]Institute of Developmental Biology and Regenerative Medicine,Southwest University,Chongqing 400715,China [2]University of Chinese Academy of Sciences(Chongqing),Chongqing Institute of Green and Intelligent Technology,Chinese Academy of Sciences,Chongqing 400714,China

出　　处：《Journal of Molecular Cell Biology》2022年第12期63-66,共4页分子细胞生物学报（英文版）

基　　金：supported by the National Natural Science Foundation of China(32270859,32000576,and32192400);the National Key R&D Program of China(2021YFA0805000);the Natural ScienceeFoundationofChongqing(cstc2020jcyj-msxmX0882).

摘　　要：Transgenesis, which inserts exogenous DNA into animal genomes, is a widely used technique. Traditionally, the constructs for transgenesis are generated by step-by-step subcloning of DNA fragments, which requires multiple steps depending on the construct complexity. To overcome the limitation, advanced tools such as Gateway cloning (Hartley et al., 2000;Kwan et al., 2007), In-Fusion cloning (Sleight et al., 2010), and Gibson assembly (Gibson et al., 2009) have been developed. However, due to their ligation characteristics, no systematic method for transgenesis has been developed. ‘Golden Gate’ cloning first appeared in 2008, which is a widely used DNA assembly method (Engler et al., 2008, 2009). Here, we take zebrafish transgenesis as an example and develop a standardized system called GoldenFish, which is based on Golden Gate cloning. It can customize transgenic constructs in one step and can be applied to multiple types of transgenesis such as one gene driven by one promoter, multiple genes driven by one promoter, and multiple genes respectively driven by multiple promoters, significantly reducing working time.

关 键 词：FISH GOLDEN SYSTEM 

分 类 号：Q78[生物学—分子生物学]