Dataset Information

The iBeetle large-scale RNAi screen reveals gene functions for insect development and physiology.

ABSTRACT: Genetic screens are powerful tools to identify the genes required for a given biological process. However, for technical reasons, comprehensive screens have been restricted to very few model organisms. Therefore, although deep sequencing is revealing the genes of ever more insect species, the functional studies predominantly focus on candidate genes previously identified in Drosophila, which is biasing research towards conserved gene functions. RNAi screens in other organisms promise to reduce this bias. Here we present the results of the iBeetle screen, a large-scale, unbiased RNAi screen in the red flour beetle, Tribolium castaneum, which identifies gene functions in embryonic and postembryonic development, physiology and cell biology. The utility of Tribolium as a screening platform is demonstrated by the identification of genes involved in insect epithelial adhesion. This work transcends the restrictions of the candidate gene approach and opens fields of research not accessible in Drosophila.

SUBMITTER: Schmitt-Engel C

PROVIDER: S-EPMC4525174 | biostudies-literature | 2015 Jul

REPOSITORIES: biostudies-literature

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The iBeetle large-scale RNAi screen reveals gene functions for insect development and physiology.

Schmitt-Engel Christian C Schultheis Dorothea D Schwirz Jonas J Ströhlein Nadi N Troelenberg Nicole N Majumdar Upalparna U Dao Van Anh VA Grossmann Daniela D Richter Tobias T Tech Maike M Dönitz Jürgen J Gerischer Lizzy L Theis Mirko M Schild Inga I Trauner Jochen J Koniszewski Nikolaus D B ND Küster Elke E Kittelmann Sebastian S Hu Yonggang Y Lehmann Sabrina S Siemanowski Janna J Ulrich Julia J Panfilio Kristen A KA Schröder Reinhard R Morgenstern Burkhard B Stanke Mario M Buchhholz Frank F Frasch Manfred M Roth Siegfried S Wimmer Ernst A EA Schoppmeier Michael M Klingler Martin M Bucher Gregor G

Nature communications 20150728

Genetic screens are powerful tools to identify the genes required for a given biological process. However, for technical reasons, comprehensive screens have been restricted to very few model organisms. Therefore, although deep sequencing is revealing the genes of ever more insect species, the functional studies predominantly focus on candidate genes previously identified in Drosophila, which is biasing research towards conserved gene functions. RNAi screens in other organisms promise to reduce t ...[more]

PMID: 26215380

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Project description:Neuronal circuit development and function require proper synapse formation and maintenance. Genetic screens are one powerful method to identify the mechanisms shaping synaptic development and stability. However, genes with essential roles in non-neural tissues may be missed in traditional loss-of-function screens. In an effort to circumvent this limitation, we used neuron-specific RNAi knock down in Drosophila and assayed the formation, growth, and maintenance of the neuromuscular junction (NMJ). We examined 1970 Drosophila genes, each of which has a conserved ortholog in mammalian genomes. Knock down of 158 genes in post-mitotic neurons led to abnormalities in the neuromuscular system, including misapposition of active zone components opposite postsynaptic glutamate receptors, synaptic terminal overgrowth and undergrowth, abnormal accumulation of synaptic material within the axon, and retraction of synaptic terminals from their postsynaptic targets. Bioinformatics analysis demonstrates that genes with overlapping annotated function are enriched within the hits for each phenotype, suggesting that the shared biological function is important for that aspect of synaptic development. For example, genes for proteasome subunits and mitotic spindle organizers are enriched among the genes whose knock down leads to defects in synaptic apposition and NMJ stability. Such genes play essential roles in all cells, however the use of tissue- and temporally-restricted RNAi indicates that the proteasome and mitotic spindle organizers participate in discrete aspects of synaptic development. In addition to identifying functional classes of genes shaping synaptic development, this screen also identifies candidate genes whose role at the synapse can be validated by traditional loss-of-function analysis. We present one such example, the dynein-interacting protein NudE, and demonstrate that it is required for proper axonal transport and synaptic maintenance. Thus, this screen has identified both functional classes of genes as well as individual candidate genes that are critical for synaptic development and will be a useful resource for subsequent mechanistic analysis of synapse formation and maintenance.

Dataset Information

The iBeetle large-scale RNAi screen reveals gene functions for insect development and physiology.

Publications

The iBeetle large-scale RNAi screen reveals gene functions for insect development and physiology.

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