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当前位置:首页  »  实验视频  »  Developmental Biology  »  Microinjection of mRNA and Morpholino Antisense Oligonucleotides in Zebrafish Embryos.

  • Microinjection of mRNA and Morpholino Antisense Oligonucleotides in Zebrafish Embryos.


    时长:未知

    类型:Developmental Biology

    时间:2009

    国家:美国

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    播放次数:加载中

    包包:20免费充值包包

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播放地址1: FLV文件(说明: 无需安装任何插件,即可快速播放 )

内容介绍

    Shiaulou Yuan, Zhaoxia SunDepartment of Genetics, Yale University School of Medicine An essential tool for investigating the role of a gene during development is the ability to perform gene knockdown, overexpression, and misexpression studies. In zebrafish (Danio rerio), microinjection of RNA, DNA, proteins, antisense oligonucleotides and other small molecules into the developing embryo provides researchers a quick and robust assay for exploring gene function in vivo. In this video-article, we will demonstrate how to prepare and microinject in vitro synthesized EGFP mRNA and a translational-blocking morpholino oligo against pkd2, a gene associated with autosomal dominant polycystic kidney disease (ADPKD), into 1-cell stage zebrafish embryos. We will then analyze the success of the mRNA and morpholino microinjections by verifying GFP expression and phenotype analysis. Broad applications of this technique include generating transgenic animals and germ-line chimeras, cell-fate mapping and gene screening. Herein we describe a protocol for overexpression of EGFP and knockdown of pkd2 by mRNA and morpholino oligonucleotide injection.

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