| Chapter title |
Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization.
|
|---|---|
| Chapter number | 24 |
| Book title |
Functional Genomics
|
| Published in |
Methods in molecular biology, November 2011
|
| DOI | 10.1007/978-1-61779-424-7_24 |
| Pubmed ID | |
| Book ISBNs |
978-1-61779-423-0, 978-1-61779-424-7
|
| Authors |
William Ka Fai Tse, Yun-Jin Jiang, Tse, William Ka Fai, Jiang, Yun-Jin |
| Editors |
Michael Kaufmann, Claudia Klinger |
| Abstract |
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. |
X Demographics
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|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 6 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Other | 1 | 17% |
| Student > Bachelor | 1 | 17% |
| Student > Ph. D. Student | 1 | 17% |
| Researcher | 1 | 17% |
| Professor > Associate Professor | 1 | 17% |
| Other | 0 | 0% |
| Unknown | 1 | 17% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 3 | 50% |
| Biochemistry, Genetics and Molecular Biology | 1 | 17% |
| Unknown | 2 | 33% |