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Generation of BAC Transgenic Mice for Functional Analysis of Neural Circuits

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Neural Tracing Methods

Part of the book series: Neuromethods ((NM,volume 92))

Abstract

Functional analysis of neural circuits in the living brain is an exceptional challenge that has been greatly advanced in the modern molecular genetics era by the development of elegant techniques for marking and manipulating genetically defined neuronal subsets. Technologies of this nature are enabling neuroscientists to probe the causal role of specific neuronal populations in sensory information processing, complex animal behaviors, and brain dysfunction in a myriad of neurological disorders. Gains in these areas have been especially catalyzed by the development and expansion of optogenetics, specifically, the use of channelrhodopsin-2 (ChR2) for inducible and reversible control of neuronal firing with blue light. Here we provide detailed methods based on our recent success in developing BAC transgenic mouse lines with functional ChR2 expression in diverse genetically-defined neuronal subsets. In principle this BAC transgenic strategy can be implemented to express other transgenes of interest or can be further applied to achieve stable transgenic expression of ChR2 and other microbial opsin variants in additional diverse cell types of the mammalian nervous system.

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Acknowledgments

We wish to acknowledge Dr. Bernd Gloss for providing the iTV1 plasmid and for significant contributions on the BAC recombineering and electroelution procedures described in this work. We thank Dr. Karl Deisseroth for providing the original ChR2(H134R)-EYFP plasmid. The EL250 strain used for BAC recombineering was generously provided by Dr. Neal Copeland. This work was supported in part by an American Recovery and Reinvestment Act grant from the US National Institute of Mental Health to G.F. (RC1-MH088434), a National Alliance for Research on Schizophrenia and Depression: The Brain and Behavior Research Foundation Young Investigator Award to J.T.T., and US National Institutes of Health Ruth L. Kirschstein National Research Service Awards to J.T.T. (F32-MH084460).

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Authors and Affiliations

  1. Department of Brain and Cognitive Sciences, McGovern Institute for Brain Research, Massachusetts Institute of Technology, Building 46-3143A, 43 Vassar St., Cambridge, MA, 02139, USA

    Jonathan T. Ting Ph.D. & Guoping Feng

Authors
  1. Jonathan T. Ting Ph.D.
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  2. Guoping Feng
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Corresponding author

Correspondence to Jonathan T. Ting Ph.D. .

Editor information

Editors and Affiliations

  1. Baylor College of Medicine, Houston, Texas, USA

    Benjamin R. Arenkiel

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Ting, J.T., Feng, G. (2015). Generation of BAC Transgenic Mice for Functional Analysis of Neural Circuits. In: Arenkiel, B. (eds) Neural Tracing Methods. Neuromethods, vol 92. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1963-5_9

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  • DOI: https://doi.org/10.1007/978-1-4939-1963-5_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1962-8

  • Online ISBN: 978-1-4939-1963-5

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