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An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in <em>Drosophila</em> by Genome Editing

Binary transcription systems are powerful genetic tools widely used for visualizing and manipulating cell fate and gene expression in specific groups of ...

University of Maryland

Fibroblast growth factors (FGF) are essential signaling proteins that regulate diverse cellular functions in developmental and metabolic processes. In Drosophila, the FGF homolog, branchless (bnl) is expressed in a dynamic and spatiotemporally restricted pattern to induce branching morphogenesis of the trachea, which expresses the Bnl-receptor, breathless (btl). Here we have developed a new strategy to determine bnl- expressing cells and study their interactions with the btl-expressing cells in the range of tissue patterning during Drosophila development. To enable targeted gene expression specifically in the bnl expressing cells, a new LexA based bnl enhancer trap line was generated using CRISPR/Cas9 based genome editing. Analyses of the spatiotemporal expression of the reporter in various embryonic stages, larval or adult tissues and in metabolic hypoxia, confirmed its target specificity and versatility. With this tool, new bnl expressing cells, their unique organization and functional interactions with the btl-expressing cells were uncovered in a larval tracheoblast niche in the leg imaginal discs, in larval photoreceptors of the developing retina, and in the embryonic central nervous system. The targeted expression system also facilitated live imaging of simultaneously labeled Bnl sources and tracheal cells, which revealed a unique morphogenetic movement of the embryonic bnl- source. Migration of bnl- expressing cells may create a dynamic spatiotemporal pattern of the signal source necessary for the directional growth of the tracheal branch. The genetic tool and the comprehensive profile of expression, organization, and activity of various types of bnl-expressing cells described in this study provided us with an important foundation for future research investigating the mechanisms underlying Bnl signaling in tissue morphogenesis.

Unique patterns of organization and migration of FGF-expressing cells during Drosophila morphogenesis.

Flow injection mass spectrometry (FIMS) combined with chemometrics was evaluated for rapidly detecting economically motivated adulteration (EMA) of milk. Twenty-two pure milk and thirty-five counterparts adulterated with soybean, pea, and whey protein isolates at 0.5, 1, 3, 5, and 10% (w/w) levels were analyzed. The principal component analysis (PCA), partial least-squares-discriminant analysis (PLS-DA), and support vector machine (SVM) classification models indicated that the adulterated milks could successfully be classified from the pure milks. FIMS combined with chemometrics might be an effective method to detect possible EMA in milk.

Lijuan Du

Department of Cell Biology and Molecular Genetics

University of Maryland

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

Lijuan Du

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Cell Biology and Molecular Genetics

University of Maryland

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

Department of Cell Biology and Molecular Genetics