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用于作为模式遗传生物的青鳉胚胎显微注射。

Microinjection of medaka embryos for use as a model genetic organism.

作者信息

Porazinski Sean R, Wang Huijia, Furutani-Seiki Makoto

机构信息

Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath.

出版信息

J Vis Exp. 2010 Dec 22(46):1937. doi: 10.3791/1937.

Abstract

In this video, we demonstrate the technique of microinjection into one-cell stage medaka embryos. Medaka is a small egg-laying freshwater fish that allows both genetic and embryological analyses and is one of the vertebrate model organisms in which genome-wide phenotype-driven mutant screens were carried out (1), as in zebrafish and the mouse. Divergence of functional overlap of related genes between medaka and zebrafish allows identification of novel phenotypes that are unidentifiable in a single species (2), thus medaka and zebrafish are complementary for genetic dissection of vertebrate genome functions. To take advantage of medaka fish whose embryos are transparent and develop externally, microinjection is an essential technique to inject cell-tracers for labeling cells, mRNAs or anti-sense oligonucleotides for over-expressing and knocking-down genes of interest, and DNAs for making transgenic lines.

摘要

在本视频中,我们展示了对单细胞期青鳉胚胎进行显微注射的技术。青鳉是一种小型的产卵淡水鱼,可用于遗传和胚胎学分析,并且是进行全基因组表型驱动突变体筛选的脊椎动物模式生物之一(1),如同斑马鱼和小鼠。青鳉和斑马鱼相关基因功能重叠的差异使得能够鉴定出在单一物种中无法识别的新表型(2),因此青鳉和斑马鱼在脊椎动物基因组功能的遗传剖析方面具有互补性。为了利用胚胎透明且在体外发育的青鳉鱼,显微注射是一种必不可少的技术,可用于注射细胞追踪剂以标记细胞、注射mRNA或反义寡核苷酸以过表达和敲低感兴趣的基因,以及注射DNA以构建转基因品系。

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本文引用的文献

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