Dreadlocks Dock is necessary to regulate growth of the germline ring canals in the developing Drosophila melanogaster egg chamber Olivia Crowe Click here to view. The Ovhts polyprotein is cleaved to produce fusome and ring canal proteins required for Drosophila oogenesis. For Kr , the main defect is the absence of the A2 segment arrowhead , which is a smaller gap than seen in classic mutant embryos. Citations Publications citing this paper.

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

Timing aside, the two drivers led to similar embryonic phenotypes and were used interchangeably in this study. The system appears especially effective at depleting genes required during mid-embryogenesis after gastrulation 4—5 hr after egg laying.

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The torso pathway in Drosophila: Functions and mechanisms of receptor tyrosine kinase Torso signaling: 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. The decapentaplegic gene is required for dorsal-ventral patterning of the Drosophila embryo. National Center for Biotechnology InformationU. Together with the data from dpp and the heterozygous mutants, these results suggest that the poor knockdown mwternal early zygotic genes stems from our inability to deliver enough shRNAs at the appropriate time.

Showing of 13 references. Abstract In a developing Drosophila melanogaster embryo, mRNAs have a maternal origin, a zygotic origin, or both. The Ovhts polyprotein is cleaved to produce fusome and ring canal proteins required for Drosophila oogenesis.


The time maternzl measure positional information: Zygotic phenotypes revealed by the expression of zygotic shRNAs by maternally loaded Gal4 protein. Images were acquired by laser scanning microscopy with two-photon excitation at nm Luengo Hendriks et al. 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.

WT data were downloaded from http: NGT 40 ; P Gal Interestingly, maternally gsl4 Gal4 protein, in combination with different copy numbers and delivery methods of UAS-shRNAscan be used to knock down zygotic transcripts in certain time windows and reveal distinct and gall4 phenotypes of pleiotropic genes.

These data suggest that shRNAs driven by mat-GAL4 are very effective at depleting the relevant transcripts in the female germline. The D-raf example illustrates how different embryonic phenotypes can be observed depending on the level of either maternal or zygotic gene activity present at a specific developmental stage. 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.

Depleting gene activities in early Drosophila embryos with the “maternal-Gal4-shRNA” system.

Maternally deposited shRNAs can deplete early zygotic transcripts only modestly, in most cases not enough to reveal a mategnal red acting on cyan. Click here to view.

The maternal Gal4 driver blue activates shRNAs redwhich deplete target transcripts green. Further, the technique will be useful for the analysis of regulatory network architecture and cis -regulatory element reporter constructs. References Publications referenced by this paper.

To test whether shRNAs would be more effective when expressed in the backbone of a miRNA normally expressed during late oogenesis and embryogenesis, we generated transgenic lines targeting the otuNotch NbcdKrgtwgand armadillo arm genes in the backbone of miR and miRaas both had been shown previously to be some of the most stable miRNAs present in unfertilized embryos Votruba This fraction is consistent with the quarter of embryos without a UAS-shRNA-N transgene surviving and reminiscent of the previously reported paternal rescue of the Notch maternal effect phenotype Lehmann et al.


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Received Aug 13; Accepted Oct A number of stable maternally deposited mRNAs have been identified by Votruba Results and Discussion shRNAs expressed in the female germline effectively knock down Mat genes To extend our previous finding that shRNAs expressed during oogenesis effectively knockdown maternally deposited transcripts, we tested a number of UAS-shRNA lines targeting various Mat genes.

All phenotypes resemble strong maternwl alleles.

This phenotype is similar across embryos with zero, one, or two copies of the UAS-shRNA-hb transgene, indicating that zygotically expressed shRNAs do not contribute to this phenotype see legend Figure 4.

This difference is useful, as in some cases early oogenesis defects that can be detected with MTD-Gal4 can be bypassed katernal mat-tub-Gal4thus allowing the production of eggs D.