利用TALEN打靶技术建立果蝇心脏发育候选基因Yippee突变品系  

Construction of Mutation Lines of Heart Developmental Candidate Gene Yippee with Gene Targeting Method TALENs

作　　者：蔡哲彦[1] 万永奇[1] 唐旻[1] 曾群[1] 朱莎莎[1] 廖四芳[1] 吴秀山[1] 袁婺洲[1] 

机构地区：[1]湖南师范大学蛋白质化学及鱼类发育生物学教育部重点实验室心脏发育研究中心,湖南长沙410081

出　　处：《激光生物学报》2013年第4期348-353,共6页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(81170229)

摘　　要：研究工作者们一直致力于寻找一种在模式生物中进行精确基因修饰的方法。近期基于TALENs的基因打靶方法受到广泛的注意,并在包括果蝇、斑马鱼、小鼠及人类多潜能细胞的多物种中取得成功。在这里,我主要介绍基于TALENs的基因修饰在果蝇模型中的应用以及果蝇心脏发育候选基因Yippee的敲除。首先,我们设计并拼接好了Yippee基因的TALENs靶位点序列,连接在核酸酶Xmn I的催化区域。在核酸酶的作用下产生双链的断裂,随后在非同源末端连接物修复系统(NHEJ)的修复下导致Yippee基因的消除或部分缺失。经过检测,得到Yippee基因的TALENs打靶效率约为9%。Researchers have long been committed to develop a strategy for precise genomic modification in model organ- ism. Recently, TALENs for genetic modification has been attracted much attention and achieved satisfactory results in many species including Drosophila, zebrafish, rats and human pluripotent cells. Here we described the application of TALENs for genetic modification in Drosophila and disrupted the Drosophila endogenous gene Yippee. First, we designed and assembled transcriptional activator like effectors （TALEs） to exon 1 of Yippee, linked to the catalytic domain of nu- clease Fok I ~ The site-directed nucleases generated double strand breaks, then were repaired by nonhomologous end- joining（NHEJ） repair system resulting in edits or small deletions within endogenous Drosophila Yippee gene at efficien- cies of up to 9%.

关 键 词：果蝇 Yippee基因 基因打靶 TALENs 

分 类 号：Q344.12[生物学—遗传学]