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Genetic engineering for in vivo optical interrogation of neuronal responses to cell type-specific silencing

Firat Terzi, Johannes Knabbe, Sidney B. Cambridge
doi: https://doi.org/10.1101/2021.01.13.426508
Firat Terzi
Department of Functional Neuroanatomy, Institute of Cell Biology and Anatomy, Heidelberg University, Im Neuenheimer Feld 307, 69120 Heidelberg, Germany Tel: +49-6221-548671
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Johannes Knabbe
Department of Functional Neuroanatomy, Institute of Cell Biology and Anatomy, Heidelberg University, Im Neuenheimer Feld 307, 69120 Heidelberg, Germany Tel: +49-6221-548671
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Sidney B. Cambridge
Department of Functional Neuroanatomy, Institute of Cell Biology and Anatomy, Heidelberg University, Im Neuenheimer Feld 307, 69120 Heidelberg, Germany Tel: +49-6221-548671
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  • For correspondence: cambridge@ana.uni-heidelberg.de
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Summary

Genetic engineering of quintuple transgenic brain tissue was used to establish a low background, Cre-dependent version of the inducible Tet-On system for fast, cell type-specific transgene expression in vivo. Co-expression of a constitutive, Cre-dependent fluorescent marker selectively allowed single cell analyses before and after inducible, tet-dependent transgene expression. Here, we used this method for acute, high-resolution manipulation of neuronal activity in the living brain. Single induction of the potassium channel Kir2.1 produced cell type-specific silencing within hours that lasted for at least three days. Longitudinal in vivo imaging of spontaneous calcium transients and neuronal morphology demonstrated that prolonged silencing did not alter spine densities or synaptic input strength. Furthermore, selective induction of Kir2.1 in parvalbumin interneurons increased the activity of surrounding neurons in a distance-dependent manner. This high-resolution, inducible interference and interval imaging of individual cells (high I5, ‘HighFive’) method thus allows visualizing temporally precise, genetic perturbations of defined cells.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted January 14, 2021.
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Genetic engineering for in vivo optical interrogation of neuronal responses to cell type-specific silencing
Firat Terzi, Johannes Knabbe, Sidney B. Cambridge
bioRxiv 2021.01.13.426508; doi: https://doi.org/10.1101/2021.01.13.426508
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Genetic engineering for in vivo optical interrogation of neuronal responses to cell type-specific silencing
Firat Terzi, Johannes Knabbe, Sidney B. Cambridge
bioRxiv 2021.01.13.426508; doi: https://doi.org/10.1101/2021.01.13.426508

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