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In vivo rapid gene delivery into postmitotic neocortical neurons using iontoporation

Nature Protocols volume 10, pages 25–32 (2015)Cite this article

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Abstract

This protocol describes a method for directing the expression of genes of interest into postmitotic neocortical neurons in vivo. Microinjection of a DNA plasmid-amphiphilic molecule mix into the neocortex followed by delivery of an ad hoc electric pulse protocol during the first few days of life in mice allows rapid, focal and efficient expression of genes in postmitotic neurons. Compared with other gene delivery techniques such as in utero electroporation and viral infection, this method allows rapid (12 h), focal (50–200 μm), mosaic-like (50 to several hundred neurons) targeting of postmitotic neurons within existing circuits. This 'iontoporation' protocol, which can be completed within ∼20 min per mouse, allows straightforward assessment of genetic constructs in postmitotic cortical neurons and subsequent genetic, histological and physiological investigations of gene function.

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Figure 1: Schematic summary of the iontoporation protocol.
Figure 2: Targeting postmitotic neurons with iontoporation.
Figure 3: Defining the stereotaxic coordinates of the target site.

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Acknowledgements

We thank the members of our laboratory for helpful discussions. We also thank A. Benoit and F. Smets for technical assistance, L. Frangeul for help with the manuscript and L. Telley for photomicrograph image acquisition. Work in the Jabaudon laboratory is supported by the Swiss National Science Foundation, the Leenaards Foundation, the Brain and Behavior Foundation and the Synapsy Foundation.

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Author notes

  1. Andres De la Rossa

    Present address: Present address: Department of Cell Biology, University of Geneva, Geneva, Switzerland.,

Authors and Affiliations

  1. Department of Basic Neurosciences, University of Geneva, Geneva, Switzerland

    Andres De la Rossa & Denis Jabaudon

  2. Clinic of Neurology, Geneva University Hospital, Geneva, Switzerland

    Denis Jabaudon

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  1. Andres De la Rossa
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  2. Denis Jabaudon
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Contributions

A.D.l.R. and D.J. developed the protocol and wrote the manuscript; A.D.l.R. performed the experiments.

Corresponding author

Correspondence to Denis Jabaudon.

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The authors declare no competing financial interests.

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De la Rossa, A., Jabaudon, D. In vivo rapid gene delivery into postmitotic neocortical neurons using iontoporation. Nat Protoc 10, 25–32 (2015). https://doi.org/10.1038/nprot.2015.001

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