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Gene Silencing by Injection and Electroporation of dsRNA in Avian Embryos


Mauti, O; Baeriswyl, T; Stoeckli, E T (2008). Gene Silencing by Injection and Electroporation of dsRNA in Avian Embryos. Cold Spring Harbor Protocols:5094.

Abstract

In ovo RNA interference (RNAi) is a method for silencing a gene of interest using a combination of in ovo injection and electroporation in avian embryos. Here we describe gene silencing in the developing spinal cord, but the procedure can easily be adapted to other parts of the nervous system. Double-stranded RNA (dsRNA) derived from the gene of interest is injected into the developing spinal cord of the chicken embryo, and is followed by electroporation to allow for the uptake of the dsRNA. With this method, temporal as well as spatial control of gene silencing is possible. The time point of injection should be chosen according to the expression profile of the gene or the half-life of the protein. Proteins with slow turnover may require RNAi at earlier stages, ideally before the onset of gene expression. The electroporation parameters can be adjusted such that only a specific population of neurons is targeted in the spinal cord.

In ovo RNA interference (RNAi) is a method for silencing a gene of interest using a combination of in ovo injection and electroporation in avian embryos. Here we describe gene silencing in the developing spinal cord, but the procedure can easily be adapted to other parts of the nervous system. Double-stranded RNA (dsRNA) derived from the gene of interest is injected into the developing spinal cord of the chicken embryo, and is followed by electroporation to allow for the uptake of the dsRNA. With this method, temporal as well as spatial control of gene silencing is possible. The time point of injection should be chosen according to the expression profile of the gene or the half-life of the protein. Proteins with slow turnover may require RNAi at earlier stages, ideally before the onset of gene expression. The electroporation parameters can be adjusted such that only a specific population of neurons is targeted in the spinal cord.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Institute of Molecular Life Sciences
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:26 November 2008
Deposited On:23 Jan 2009 15:37
Last Modified:05 Apr 2016 12:48
Publisher:Cold Spring Harbor Press
ISSN:1559-6095
Publisher DOI:10.1101/pdb.prot5094
Related URLs:http://cshprotocols.cshlp.org/cgi/content/abstract/2008/13/pdb.prot5094?maxtoshow=&HITS&hits&RESULTFORMAT=&fulltext=stoeckli&searchid=1&FIRSTINDEX=0&resourcetype=HWCIT
Permanent URL: http://doi.org/10.5167/uzh-9753

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