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In vivo electroporation in the embryonic mouse central nervous system

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Abstract

This protocol describes a basic method for in vivo electroporation in the nervous system of embryonic mice. Delivery of electric pulses following microinjection of DNA into the brain ventricle or the spinal cord central canal enables efficient transfection of genes into the nervous system. Transfection is facilitated by forceps-type electrodes, which hold the uterus and/or the yolk sac containing the embryo. More than ten embryos in a single pregnant mouse can be operated on within 30 min. More than 90% of operated embryos survive and more than 90% of these survivors express the transfected genes appropriately. Gene expression in neurons persists for a long time, even at postnatal stages, after electroporation. Thus, this method could be used to analyze roles of genes not only in embryonic development but also in higher order function of the nervous system, such as learning.

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Figure 1: A pregnant mouse on the operating board.
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Figure 4: DNA injection into the in utero embryo.
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Figure 8: Neurons labeled with EYFP.
Figure 9: Long-term expression of the mitochondrial-targeted DsRed protein.

Change history

  • 29 December 2006

    In the version of this article initially published online, several instances of the phrase in vivo were incorrectly substituted with in ovo. When the article was first corrected, not all instances were changed appropriately. These errors have been corrected in all versions of the article. In addition, on p. 1557 the first line of the Troubleshooting section should read “Steps 6A(i)–(iv) and 6B(i)–(iv)…” rather than “Steps 6A(i)–(iv) and 6B(ix)–(xii)…”

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Acknowledgements

I thank J. Kutsuna, M. Tanaka and Y. Yamamoto for their help and Dr. D. Kawauchi for critical reading of the manuscript. This work was supported by Brain Science Foundation and grants from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Correspondence to Tetsuichiro Saito.

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Saito, T. In vivo electroporation in the embryonic mouse central nervous system. Nat Protoc 1, 1552–1558 (2006). https://doi.org/10.1038/nprot.2006.276

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