A genome engineered tool set for Drosophila TGF-β/BMP signaling studies.

IF 3.7 2区生物学 Q1 DEVELOPMENTAL BIOLOGY Development Pub Date : 2024-11-04 DOI:10.1242/dev.204222

Clara-Maria Ell, Abu Safyan, Mrinal Chayengia, Manuela M M Kustermann, Jennifer Lorenz, Melanie Schächtle, George Pyrowolakis

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Abstract

Ligands of the TGF-β/BMP superfamily are critically involved in the regulation of growth, patterning and organogenesis and can act as long-range morphogens. Essential for understanding TGF-β/BMP signaling dynamics and regulation are tools that allow monitoring and manipulating pathway components expressed at physiological levels and endogenous spatiotemporal patterns. We used genome engineering to generate a comprehensive library of endogenously epitope- or fluorescently-tagged versions of receptors, co-receptors, transcription factors and key feedback regulators of the Drosophila BMP and Activin signaling pathways. We demonstrate that the generated alleles are biologically active and can be utilized for assessing tissue and subcellular distribution of the corresponding proteins. Further, we show that the genomic platforms can be used for in locus structure-function and cis-regulatory analyses. Finally, we present a complementary set of protein binder-based tools, which allow visualization as well as manipulation of the stability and subcellular localization of epitope-tagged proteins, providing new tools for the analysis of BMP signaling and beyond.

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用于果蝇 TGF-β/BMP 信号研究的基因组工程工具集。

TGF-β/BMP 超家族配体在生长、模式化和器官形成的调控中发挥着关键作用，并可作为长程形态发生因子。要了解 TGF-β/BMP 信号动态和调控，关键是要有能监测和操纵以生理水平和内源性时空模式表达的通路成分的工具。我们利用基因组工程生成了一个全面的内源表位或荧光标记的受体、共受体、转录因子以及果蝇 BMP 和 Activin 信号通路的关键反馈调节因子库。我们证明所生成的等位基因具有生物活性，可用于评估相应蛋白质的组织和亚细胞分布。此外，我们还展示了基因组平台可用于基因座结构-功能和顺式调控分析。最后，我们还介绍了一套基于蛋白质粘合剂的补充工具，它可以对表位标记蛋白质的稳定性和亚细胞定位进行可视化操作，为 BMP 信号转导及其他分析提供了新的工具。

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来源期刊

Development 生物-发育生物学

CiteScore

6.70

自引率

4.30%

发文量

433

审稿时长

3 months

期刊介绍： Development’s scope covers all aspects of plant and animal development, including stem cell biology and regeneration. The single most important criterion for acceptance in Development is scientific excellence. Research papers (articles and reports) should therefore pose and test a significant hypothesis or address a significant question, and should provide novel perspectives that advance our understanding of development. We also encourage submission of papers that use computational methods or mathematical models to obtain significant new insights into developmental biology topics. Manuscripts that are descriptive in nature will be considered only when they lay important groundwork for a field and/or provide novel resources for understanding developmental processes of broad interest to the community. Development includes a Techniques and Resources section for the publication of new methods, datasets, and other types of resources. Papers describing new techniques should include a proof-of-principle demonstration that the technique is valuable to the developmental biology community; they need not include in-depth follow-up analysis. The technique must be described in sufficient detail to be easily replicated by other investigators. Development will also consider protocol-type papers of exceptional interest to the community. We welcome submission of Resource papers, for example those reporting new databases, systems-level datasets, or genetic resources of major value to the developmental biology community. For all papers, the data or resource described must be made available to the community with minimal restrictions upon publication. To aid navigability, Development has dedicated sections of the journal to stem cells & regeneration and to human development. The criteria for acceptance into these sections is identical to those outlined above. Authors and editors are encouraged to nominate appropriate manuscripts for inclusion in one of these sections.