The Fly House provides research support for those scientists at IMBA using the fruit fly Drosophila melanogaster as a genetic model system. Our services include the generation of transgenic fly lines, gene targeting to generate knock-out or knock-in mutants, and large-scale in vivo RNAi screens.
A fundamental service of the Fly House is the microinjection of Drosophila embryos in order to generate stable transgenic fly lines. We routinely inject DNA constructs, including large BAC clones, into a range of commonly used host strains as well as attP landing site stocks for phiC31-mediated targeted integration. Over the course of a year, around 250 different constructs get injected at the facility.
Genome engineering in Drosophila via traditional homologous recombination is a time-consuming and labour-intensive process. Recent advances in the development and application of programmable nucleases significantly enhanced gene-targeting rates. After setting up a versatile and efficient genome engineering platform using the CRISPR/Cas9 technology, we apply this strategy to generate frame-shift mutations (indels), defined point mutations, precise deletions as well as to add epitope tags to endogenous genes via homology-directed repair (HDR). Rather than screening half a million flies required with the conventional method, we are now able to isolate targeted mutations from 100 flies in a process that takes two months.
The first genome-wide collection of inducible RNAi lines, available through the Vienna Drosophila Resource Center (VDRC), has revolutionized loss-of-function genetic screens in Drosophila. It is now possible to systematically investigate the function of each gene in a tissue of interest. We support scientists conducting large-scale in vivo RNAi screens as well as with subsequent follow up experiments to validate their candidate genes.
Efficient CRISPR/Cas9 plasmids for rapid and versatile genome editing in Drosophila.
Gokcezade J, Sienski G, Duchek P.
G3 (Bethesda). 2014 Sep 17;4(11):2279-82.
BIOSAFETY. Safeguarding gene drive experiments in the laboratory.
Akbari OS, Bellen HJ, Bier E, Bullock SL, Burt A, Church GM, Cook KR, Duchek P, Edwards OR, Esvelt KM, Gantz VM, Golic KG, Gratz SJ, Harrison MM, Hayes KR, James AA, Kaufman TC, Knoblich J, Malik HS, Matthews KA, O'Connor-Giles KM, Parks AL, Perrimon N, Port F, Russell S, Ueda R, Wildonger J.
Science. 2015 Aug 28;349(6251):927-9.
A Combination of CRISPR/Cas9 and Standardized RNAi as a Versatile Platform for the Characterization of Gene Function.
Wissel S, Kieser A, Yasugi T, Duchek P, Roitinger E, Gokcezade J, Steinmann V, Gaul U, Mechtler K, Förstemann K, Knoblich JA, Neumüller RA.
G3 (Bethesda). 2016 Aug 9;6(8):2467-78.
Linear ubiquitination by LUBEL has a role in Drosophila heat stress response.
Asaoka T, Almagro J, Ehrhardt C, Tsai I, Schleiffer A, Deszcz L, Junttila S, Ringrose L, Mechtler K, Kavirayani A, Gyenesei A, Hofmann K, Duchek P, Rittinger K, Ikeda F.
EMBO Rep. 2016 Nov;17(11):1624-1640.