This phenotype strongly resembles that of embryos that lack maternal hb and have reduced zygotic hb Simpson-Brose et al. Our ability to detect a cuticle phenotype for dpp most likely reflects the haploinsufficiency associated with this gene Spencer et al. All oligos used are listed in supporting information, Table S1. This second class resembles embryos that have substantially reduced zygotic expression of anterior hb Wimmer et al. Embryos that do not receive a paternal copy show poor cuticle development, reflecting the role of D-raf downstream of another RTK, the epidermal growth factor receptor EGFR , which is required for proper epidermal differentiation.

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Depleting Gene Activities in Early Drosophila Embryos with the “Maternal-Gal4–shRNA” System

These data suggest that shRNAs driven by mat-GAL4 are very effective at depleting the relevant transcripts in the female germline. Click here to view.

We looked for phenotypes in sensitized backgrounds for the following genes: Zygotic le – thals with speci fi c maternal effect phenotypes in Drosophila melanogaster. Luengo HendriksSoile V. Finally, all mutant alleles were obtained from the Bloomington Drosophila Stock Center: Early patterning genes escape knockdown because maternally loaded shRNAs lose efficacy over time, and zygotically expressed shRNAs are activated too late.

A conserved developmental patterning network produces quantitatively different output in multiple species of Drosophila. Thus, varying the copy number of zygotic UAS-shRNA transgenes provides a useful way to generate phenotypic series and uncover when pleiotropic genes are used in development.

FlyBase Recombinant Construct Report: P{matα4-GAL-VP16}

However, the efficacy of additional shRNAs was unmasked by generating heterozygous mutant zygotic backgrounds. Maternally deposited shRNAs can deplete early zygotic transcripts only modestly, in most cases not enough to reveal a phenotype red maternsl on cyan. The difference between MTD-Gal4 and mat-tub-Gal4 driver lines is that mat-tub-Gal4 does not drive expression during early oogenesis in the germarium. Materials and Methods Drosophila strains Two different maternal Gal4 drivers maternal-Gal4 were used: Together, these data suggest how, for genes expressed after gastrulation, maternal Gal4 can activate zygotically delivered shRNAs to strongly deplete target transcripts.


In addition to lacking T2 and T3, these embryos lack the A1 abdominal segment and head structures B4.

The torso pathway in Drosophila: Embryonic phenotypes associated with knockdown of Mat and Zyg genes. Supplementary Material Supporting Information: For some genes, high rates of F 1 lethality and specific embryonic phenotypes were detected when maternal-Gal4 females were crossed to UAS-shRNA males.

Maternally deposited hb mRNA is selectively translated in the anterior and degraded in gak4 regions, while zygotic hb is expressed in an anterior domain and a posterior stripe.

From This Paper Figures, tables, and topics from this paper. B2 shows the head of materbal in B1. Knowles Genome Biology Embryo length was normalized and expression boundaries were detected by finding the inflection point in the pattern.

The Ovhts polyprotein is cleaved to produce fusome and ring canal proteins required for Drosophila oogenesis. The origin of pattern and polarity in the Drosophila embryo. Krknigap ; hkbfork head fkh terminal ; eveftz pair rule ; twist twisnail sna dorsal—ventral ; wghhand en segment polarityand were able to detect clear phenotypes for shRNAs targeting Kr and twi.


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Briefly, the sytox green nuclear stain was used to automatically identify nuclei and the Cy3 signal in each nucleus was quantified Luengo Hendriks et al. Although we cannot be certain that all the UAS-shRNA lines are effective at knocking down the targeted transcripts, these results indicate that most shRNAs delivered from the mother to the embryo do not sufficiently deplete early zygotic transcripts to generate embryonic phenotypes detectable in the cuticle.

Zygotic lethal mutations with maternal effect phenotypes in Drosophila melanogaster. In addition, loading the embryo with shRNAs against early zygotic genes was effective in only a few cases dpp and hbpossibly because shRNAs are unstable see model in Figure 6.

Topics Discussed in This Paper. To interrogate the function of mRNAs that are both maternally and zygotically expressed, it is common to examine the embryonic phenotypes derived from female germline mosaics.

Support Center Support Center. However, while the roles of D-Raf and Matenral in Tor signaling have been well characterized by the analysis of their GLC phenotypes Duffy and Perrimon ; LiRl has only been implicated in Tor signaling by the ability of a rl loss-of-function mutation to suppress a gain-of-function Tor mutation Brunner et al.

Articles from Genetics are provided here courtesy of Genetics Society of America.