In ovo RNAi – a tool to study gene function in the developing nervous system

RNAi (RNA interference) describes the still not fully understood process by which the presence of double-stranded RNA in a cell blocks the formation of the gene product from the respective gene. This process was first discovered serendipitously in Caenorhabditis elegans but it is conserved throughout the animal kingdom including humans and also in plants.


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We have established in ovo RNAi, a technique to silence genes in the developing nervous system of chicken embryos that is based on RNAi and in ovo electroporation. The major advantage of this approach is the temporal and spatial control over gene function. Very often genes are actively involved in more than one process during development. Gene knockouts with classical genetic tools prevent the analysis of later events because a loss of gene function during early development will mask later events or induce early embryonic lethality.
The importance of temporal control over loss of gene function is well demonstrated in our studies identifying the morphogen shh (sonic hedgehog) as a guidance cue for postcommissural axons in the spinal cord.

 

Publications:

Bourikas, D., and Stoeckli, E.T. (2003) New tools for gene manipulation in chicken embryos Oligonucleotides, 13: 411-419   pubmed
Pekarik, V., Bourikas, D., Miglino, N., Joset, P., Preiswerk, S., and Stoeckli, E.T. (2003) Screening for gene function in chicken embryo using RNAi and electroporation Nat. Biotechnol., 21: 93-96   pubmed