FlpStop, a tool for conditional gene control in Drosophila

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Abstract

Manipulating gene function cell type-specifically is a common experimental goal in Drosophila research and has been central to studies of neural development, circuit computation, and behavior. However, current cell type-specific gene disruption techniques in flies often reduce gene activity incompletely or rely on cell division. Here we describe FlpStop, a generalizable tool for conditional gene disruption and rescue in post-mitotic cells. In proof-of-principle experiments, we manipulated apterous, a regulator of wing development. Next, we produced conditional null alleles of Glutamic acid decarboxylase 1(Gad1) and Resistant to dieldrin (Rdl), genes vital for GABAergic neurotransmission, as well as cacophony (cac) and paralytic (para), voltage-gated ion channels central to neuronal excitability. To demonstrate the utility of this approach, we manipulated cac in a specific visual interneuron type and discovered differential regulation of calcium signals across subcellular compartments. Thus, FlpStop will facilitate investigations into the interactions between genes, circuits, and computation.

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Yvette Erica Fisher

Department of Neurobiology, Harvard Medical School, Boston, United States

Competing interests
The authors declare that no competing interests exist.
Helen H Yang

Department of Neurobiology, Stanford University, Stanford, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-5140-9664
Jesse Isaacman-Beck

Department of Neurobiology, Stanford University, Stanford, United States

Competing interests
The authors declare that no competing interests exist.
Marjorie Xie

Department of Neurobiology, Stanford University, Stanford, United States

Competing interests
The authors declare that no competing interests exist.
Daryl M Gohl

University of Minnesota Genomics Center, Minneapolis, United States

Competing interests
The authors declare that no competing interests exist.
Thomas R Clandinin

Department of Neurobiology, Stanford University, Stanford, United States

For correspondence
trc@stanford.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-6277-6849

Funding

National Eye Institute (R01 EY022638)

Thomas R Clandinin

National Institute of Mental Health (U01 MH109119)

Thomas R Clandinin

National Science Foundation

Yvette Erica Fisher

Stanford University School of Medicine

Helen H Yang

National Eye Institute (F32EY020040)

Daryl M Gohl

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

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This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.