Gain- and loss-of-function in chick embryos by electroporation

Abstract

It remained very difficult to manipulate gene expression in chick embryos until the advent of in ovo electroporation which enabled the induction of both gain-of-function, and recently loss-of-function, of a gene of interest at a specific developmental stage. Gain-of-function by electroporation is so effective that it has become widely adopted in developmental studies in the chick. Recently, it became possible to induce loss-of-function by introducing an siRNA expression vector by electroporation. In this review, the methods of electroporation for gain-of-function and for loss-of-function by siRNA are discussed.

Neural induction: old problem, new findings, yet more questions

“…However, this technique does not allow one to restrict the gain or loss of gene function to a particular cell group at a particular stage, which is a problem when the molecules of interest have multiple sequential roles (as do BMPs, FGFs and Wnts). The introduction of focal electroporation in the chick (Nakamura et al, 2004), coupled with its large size and the ease with which transplants can be performed, has enabled spatiotemporally controlled misexpression and knockdown studies that are now starting to reveal the complexity of the signalling processes that underlie neural induction. The chick also possesses a very large territory that does not contribute cells to the neural plate or to its border, a prerequisite for any assay of induction, which requires an assessment of whether cells can be diverted to a new fate or whether the normal neural territory is expanded or compressed.…”

Chicken genome: Current status and future opportunities

“…A total of 4532 fulllength cDNA clones (Caldwell et al 2004;Hubbard et al 2005), representing ∼25% of known gene predictions in chicken, can now be used in evolutionary and functional studies (available from ARK-Genomics). RNAi and transgenic technologies are now available in the chicken, which when combined with the accessible chicken embryo, makes this a powerful system for functional studies in vivo (Brown et al 2003;Nakamura et al 2004;Sang 2004;Stern 2004). The application of these tools and access to the biological information they generate is a huge and complex task.…”

Evaluation of three different promoters driving gene expression in developing chicken embryo by using in vivo electroporation

“…Compared to virus vectors, plasmid vectors are safer and easier for use in studies. Recently, the method of in vivo electroporation binding plasmid vector has become popular for studying the gain and loss of functions to exogenous gene expression, especially with respect to elucidating the development of the embryonic central nervous system (Muramatsu et al, 1997;Katahira and Nakamura, 2003;Chesnutt and Niswander, 2004;Nakamura et al, 2004;Das et al, 2006;Sauka-Spengler and Barembaum, 2008;Lin et al, 2011). In the same embryo after electroporation, the electroporated side represents the experimental area, while the other side serves as the control.…”