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Besides testing trans-heterozygous flies for lethality, we also tested their fertility, as some of our mutants were not lethal.
To circumvent this lethality, we also generated a new Gal4 driver, IFM-GAL4, in which GAL4 expression is driven specifically in the IFM, using regulatory sequences from the flightin gene.
To exclude that the heat shock caused the lethality, we also subjected embryos from the same clutch that were not double transgenic to heat shock; all embryos appeared normal, indicating that the lethality that was observed in double transgenic animals was, indeed, owing to the overexpression of EGFP-HRAS-G12V.
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We also recorded lethality after the nestmate interaction assays (see below).
While we also observed embryonic lethality around E7, the ratio of pups born for the +/+, +/− and −/− genotype was 25∶55∶0 for our model eliminating exon 2, whereas the model disrupting exon 3 [34] reported a ratio of 414∶515∶0.
DOI: http://dx.doi.org/10.7554/eLife.03270.012 We also found synthetic lethality in clonal cells with loss of H+ efflux and oncogene expression.
We also scored embryonic lethality for the F2 self-progeny produced by heterozygous F1 ts/+ worms raised at the permissive temperature of 15°, after shifting the F2 self-progeny to the restrictive temperature as L4s.
Moreover, synthetic lethality is also observed with Kar3Δ deletion.
Lethality was also observed for the N-terminal truncation and W557S with the muscle-specific drivers Mef2 and how24B, suggesting that the lethality associated with ubiquitous expression was likely due to expression in muscle.
This lethality was also enhanced by heat shock, thus precluding the analysis of the role of p53 in the lethality caused by Mnn1 mis-expression and stress (our unpublished results).
Furthermore, synthetic lethality predictions also aided in the prediction of superadditive drug combinations.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com