The following D. melanogaster stocks were used: UAS-RNAi lines were sourced from Vienna Drosophila Resource Center (VDRC) and stock numbers are as listed in Additional file 1 : Table S3, w1118 (BL5905), and Hand-GFP; 4 × Hand-GAL4/CyO-YFP was obtained from [42 (link)]. All lines were maintained and crosses performed at 25 °C on standard media.
Uas rnai lines
Manufactured by Vienna Drosophila Resource Center
UAS-RNAi lines are genetic tools used to study gene function in Drosophila melanogaster, the fruit fly. These lines contain inducible RNA interference (RNAi) constructs that can be used to knock down the expression of specific target genes. The UAS-RNAi lines allow for spatially and temporally controlled gene silencing in Drosophila.
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4 protocols using uas rnai lines
1
Drosophila Genetic Manipulation Protocol
2
Drosophila RNAi and Mutant Loss-of-Function Lines
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Publicly available RNAi [57 (link), 60 (link)] or mutant [6 (link), 7 (link), 80 (link), 81 (link)] loss-of-function (LOF) fly lines targeting canonical translation factors were obtained from the Bloomington Drosophila Stock Center (BDSC). Additional UAS-RNAi lines targeting eIF4B and eIF4H1 were obtained from Vienna Drosophila Resource Center (VDRC) [18 (link)].
3
Drosophila Neuronal Phenotype Assay
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Flies were maintained on standard cornmeal medium at 25 °C. parabss were gifts from Dr. Kevin O’Dell (University of Glasgow). The elav-GAL4C155 (stock no. 458) was obtained from Bloomington and UAS-RNAi lines were obtained from the Vienna Drosophila Resource Center. Parabss;GAL4Cha(19B)was derived by crossing parabss with GAL4Cha(19B) (a gift from Dr. Paul Salvaterra, City of Hope, USA).
4
RNAi Knockdown of Region C2 Genes in Drosophila
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We conducted RNAi knockdown of all of the genes within region C2 (with the exception CG34429 for which there was no available UAS line) in D. melanogaster using UAS-RNAi lines from Vienna Drosophila Resource Center (VDRC, www.vdrc.at ) and from Bloomington Drosophila Stock Center (Dataset S8 ). UAS males of our candidate genes were crossed to NP6333–GAL4 driver female virgins (P(GawB)PenNP6333) (45 (link)) carrying the transgene UAS-Dicer-2 P(UAS-Dcr-2.D). Crosses for the RNAi were carried out at 25 °C. The genital morphology of the male knockdowns was compared to NP6333-GAL4, UAS-Dicer, and UAS-RNAi controls. Clasper bristle number and tibia length were measured for 16 individuals of each genotype. Differences in clasper bristle number and tibia size were assessed using a one-way ANOVA followed by a Tukey’s test (Dataset S4 ). For raw phenotypic data, see Dataset S2a .
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