We support those scientists at IMBA who use the fruit fly Drosophila melanogaster or various nematode species, including C. elegans, as genetic model systems for their research. Our services include micro-injections to generate transgenic animals as well as CRISPR-mediated gene-targeting to establish knock-out or knock-in mutants. In addition, we aim to improve existing targeting methods and establish new techniques at IMBA.
We provide an injection service for fruit flies (D. melanogaster) as well as different nematode species (C. elegans, C. tropicalis, C. briggsae). For this purpose, we keep a range of attP landing site stocks for phiC31-mediated integration in Drosophila and use the split-hygR method for nematodes (Stevenson et al; G3, 2020). Scientists need to supply clean plasmid preps and will receive injected animals to screen for transgenic individuals.
The facility offers a complete CRISPR genome editing service in both flies and worms. IMBA researchers specify the desired modification (e.g. frame-shift alleles, point mutations, precise deletions, or insertions of short epitope tags as well as larger fluorescent tags) and we will determine a suitable targeting strategy, select guides and design donors, order and clone reagents, and then screen for targeted chromosomes. In the end, our customers receive stable lines of sequence-verified animals for their research projects.
We provide introductions for scientists who want to start working with our supported model organisms, offer advice for genotyping, planning of genetic crosses and larger screens, as well as introductions to our equipment.
- Two Zeiss inverted microscopes with injection set ups for Drosophila embryos or nematodes
- Needle pullers to produce injection needles (Sutter Instrument)
- Narishige needle grinder to open the glass capillaries prior to injection
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.