國家衛生研究院 NHRI:Item 3990099045/6091
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    題名: Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization
    作者: Tse, WK;Jiang, YJ
    貢獻者: Institute of Molecular and Genomic Medicine
    摘要: In order to unfold the function of genes, solely performing mRNA over-expression is not enough nowadays. Traditional protein expression experiments, such as Western blotting and immunohistochemical staining, could only provide researchers the changes of expression levels and/or location of their targets. To make a more strong and convincing statement about gene function, it is necessary to perform both "gain-of-function" and "loss-of-function" studies. Both assays can be performed easily by transfecting DNA plasmid and siRNA in cell culture system; while in zebrafish, mRNA and morpholino (MO) microinjection can serve similar purposes. It is common for the zebrafish community to carry out microinjection experiments to explore a gene function. Instead of making a single knockdown/over-expression of a gene, we foresee that more and more large-scale screens on certain protein families will be performed in the future. Here, based on our previous experience in zebrafish "loss-of-function" screening on deubiquitylating enzymes, we describe a general work flow, from morpholino designation, in situ hybridization, to data analysis, as a reference for researchers who may be interested in a similar screen.
    日期: 2012-01
    關聯: Methods in Molecular Biology. 2012 Jan;815(Pt 5):321-331.
    Link to: http://dx.doi.org/10.1007/978-1-61779-424-7_24
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000299396500024
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84555187850
    顯示於類別:[江運金] 圖書

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