Clonal and Widespread Gene Transfer by Proviral Electroporation for Analysis of Brain Laminar Formation
An essential approach to understanding the mechanisms of development is to alter a gene function/expression. In vivo electroporation has been adapted as one such technique (Muramatsu et al., 1997). It is a very useful tool to achieve a gain- and loss-of-function (by using RNAi or morpholinos) of a gene of interest (Funahashi et al., 1999; Fukuchi-Shimogori and Grove, 2001; Kos et al., 2001; Katahira and Nakamura, 2003; Sugiyama and Nakamura, 2003). The technique has allowed the altering of gene expression temporally and spatially. Pulse-labeling technique is an approach to manipulate a specific cell population temporally, depending on its birthday, as this chapter describes. This technique is more advantageous over the BrdU application, as it can reveal cell lineage; it also has the ability to manipulate a gain- and loss-of-function into specific precursor cells (Tabata and Nakajima, 2001; Sugiyama and Nakamura, 2003; Huber et al., 2008).
In spatial terms, widespread gene transfer by electroporation has provided an efficient way to unveil a new gene function on a given tissue (Nakamura and Funahashi, 2001). On the other hand, the spatial precision of gene transfer also has been addressed as each individual cell responds differently to gene expression. Thus, there has been increasing efforts to improve electrodes and create new techniques such as single-cell electroporation (Haas et al., 2001; Kitamura et al., 2008).
KeywordsOptic Tectum Neuroepithelial Cell Morpholino Oligonucleotide Tungsten Microelectrode Deep Lamina
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