FlpStop, a tool for conditional gene control in Drosophila

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.DOI: http://dx.doi.org/10.7554/eLife.22279.001

Elementary Motion Detection inDrosophila: Algorithms and Mechanisms

“…The experimental dissection of motion detection has used a wide range of techniques, but targeted molecular manipulations have rarely been utilized. We believe that such experiments can provide fundamental new insight and are now eminently tractable, especially in light of recent technical developments (Caussinus et al 2012, Dietzl et al 2007, Fisher et al 2017, Nagarkar-Jaiswal et al 2017, Xue et al 2014. More precisely, we propose cell-type-specific gene disruption of key molecular components involved in synaptic signaling or neuronal biophysics combined with a range of visual stimuli and physiological measurements of the appropriate cell type.…”

Motor circuit function is stabilized during postembryonic growth by anterograde trans-synaptic Jelly Belly - Anaplastic Lymphoma Kinase signaling

“…We term this technique BOnStop (Bxb1 dependent on-switchable stop codon). A similar, Flp recombinase dependent technique, called FlpStop, has recently been shown to be highly effective 32 .…”

An efficient CRISPR-based strategy to insert small and large fragments of DNA using short homology arms

“…SA-T2A-LexA, SA-T2A-split GAL4; Gnerer et al, 2015;Diao et al, 2015) to obtain finer tools to express transgenes in specific cell populations. SICs can also be used to generate conditional alleles of targeted genes (Flip-flop and FLPstop (Fisher et al, 2017;Nagarkar-Jaiswal et al, 2017). Finally, strategies have been developed to convert SICs through genetic crosses rather than by injection (Trojan Exons and Double Header;Nagarkar-Jaiswal et al, 2015b;Diao et al, 2015;Li-Kroeger et al, 2018).…”