By Harukazu Nakamura Phd (auth.), Harukazu Nakamura Phd (eds.)
In ovo electroporation is an epoch-making fulfillment within the examine of developmental biology. With this system, experiments will be conducted in achieve and lack of functionality in wanted tissue at any wanted degree in chick embryos. advent of a tetracycline-regulated gene expression procedure and a transposon approach has additional prolonged the possibility of the tactic, making it attainable to acquire long term expression and to show off and on a gene of curiosity. it truly is now utilized to mice, aquatic animals, or even to vegetation for the research of developmental biology and for different reasons. during this e-book, the applying of electroporation in lots of embryonic tissues and organs is brought, with a few chapters that take care of gene move in adults. Sonoporation, one other useful gizmo, utilizing ultrasonic waves rather than electrical currents, for gene move to mesenchymal tissues can also be brought.
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Extra info for Electroporation and Sonoporation in Developmental Biology
2000). Misexpression of genes in brain vesicles by in ovo electroporation. Dev Growth Differ 42, 199–201. , Miyazaki, J. (1991). Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108, 193–199. , Takahashi, Y. (2007). Stable integration and conditional expression of electroporated transgenes in chicken embryos. Dev Biol 305, 616–624. , Takahashi, Y. (2008). Notch mediates the segmental specification of angioblasts in somites and their directed migration toward the dorsal aorta in avian embryos.
To conclude, electroporation for misexpression and silencing could be versatile by appropriate setting of a suitable condition of electroporation, such as electrodes, electric parameters, plasmids, and target tissues. 5 Troubleshooting For effective transfection, appropriate conditions of electroporation should be set. When the embryo is damaged after electroporation, you must alter the conditions such as the position of electrodes, the shape of electrodes, or the electric parameters. In general, lower voltage is used when distance between the electrodes is shorter, and narrower-exposure electrodes make it possible to transfect a narrower region (Sugiyama and Nakamura, 2003).
Mr. Imada has cooperatively supplied electrodes that are suitable for targeting tissue. , Miyazaki, J. (1998). Gene transfer into muscle by electroporation in vivo. Nat Biotechnol 16, 867–870. Funahashi, J. (1997). Gene transfer to chick embryos by electroporation (Japanese). Cell Technol 16, 1666–1675. , Nakamura, H. (1999). Role of Pax-5 in the regulation of a mid-hindbrain organizer’s activity. Dev Growth Differ 41, 59–72. , Dubrulle, J. (2005). In vivo analysis of mRNA stability using the Tet-off system in the chicken embryo.