Electroporation Rnai And Morpholinos

One of the requirements for a good model system is the ability to do gain- and loss-of-function experiments. Techniques exist to perform such experiments in chicken. A number of investigators utilize electroporation to introduce exogenous DNA into the chicken embryo in ovo (Muramatsu et al., 1997). For an excellent review of electroporation techniques in ovo, see Krull (2004). Loss-of-function experiments can be conducted by introducing short, interfering RNAs (siRNAs) or morpholinos into the chick embryo.

Double-stranded RNA-mediated interference (RNAi), a naturally occurring mechanism which results in the silencing of gene expression, has become a very powerful tool for experimental gene suppression in a number of organisms. The phenotypes observed with RNAi silencing of gene expression range from knockdown to knockout (Agrawal et al., 2003). RNAi was successfully exploited in chicken (in ovo) for gene silencing (Bourikas and Stoeckli, 2003; Pekarik et al., 2003; Krull 2004; Sato et al., 2004). In addition to siRNAs, RNAi morpholinos were used in loss-of-function studies in chicken. For example, Sheng et al. (2003) used morpho-lino oligonucleotides to knock down expression of genes in the future neural plate of the chicken embryo.

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