In ovo electroporation of miRNA plasmids to silence genes in a temporally and spatially controlled manner

Wilson, Nicole H. and Stoeckli, Esther T. (2016). In ovo electroporation of miRNA plasmids to silence genes in a temporally and spatially controlled manner. In MicroRNA Technologies (pp. 161-181) New York, NY, United States: Humana Press. doi:10.1007/7657_2016_5


Author Wilson, Nicole H.
Stoeckli, Esther T.
Title of chapter In ovo electroporation of miRNA plasmids to silence genes in a temporally and spatially controlled manner
Title of book MicroRNA Technologies
Place of Publication New York, NY, United States
Publisher Humana Press
Publication Year 2016
Sub-type Research book chapter (original research)
DOI 10.1007/7657_2016_5
Open Access Status Not yet assessed
Series Neuromethods
ISBN 9781493971732
9781493971756
ISSN 1940-6045
0893-2336
Volume number 128
Chapter number 12
Start page 161
End page 181
Total pages 21
Total chapters 15
Language eng
Subjects 2800 Neuroscience
1300 Biochemistry, Genetics and Molecular Biology
3000 Pharmacology, Toxicology and Pharmaceutics
2738 Psychiatry and Mental health
Abstract/Summary The ability to spatially and temporally control gene expression during development is crucial for the elucidation of gene function in vivo. The use of RNA interference (RNAi)-based technologies in combination with oviparous animal models allows for efficient, precise gene silencing. We have developed approaches using RNAi in the chicken embryo to analyze gene function during neural tube development. Here we describe the construction of plasmids that direct the expression of one or two artificial microRNAs (miRNAs) to knock down endogenous target protein/s upon electroporation into the spinal cord. The miRNA cassette is directly linked to a fluorescent protein reporter, which allows the faithful visualization of transfected cells. Different promoters/enhancers drive transcript expression in genetically defined cell subpopulations in the neural tube. Mixing multiple RNAi vectors allows combinatorial knockdowns of two or more genes in different cell types, thus permitting the rapid analysis of complex cellular and molecular interactions.
Keyword Axon guidance
Chicken embryo
Commissural neuron
Floor plate
Gene knockdown
MicroRNA
Neural development
RNA interference
RNAi
Spinal cord
Q-Index Code B1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Book Chapter
Collections: HERDC Pre-Audit
Queensland Brain Institute Publications
 
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Created: Fri, 09 Feb 2018, 01:51:40 EST