Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, 79104 Freiburg, Germany.
CIBSS - Centre for Integrative Biological Signalling Studies, University of Freiburg, 79104 Freiburg, Germany.
Development. 2024 Nov 15;151(22). doi: 10.1242/dev.204222. Epub 2024 Nov 18.
Ligands of the TGF-β/BMP superfamily are crucially 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 at physiological expression levels and endogenous spatiotemporal patterns. We used genome engineering to generate a comprehensive library of endogenously epitope- or fluorescent-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 used for assessing tissue and subcellular distribution of the corresponding proteins. Furthermore, 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.
TGF-β/BMP 超家族的配体在调节生长、模式形成和器官发生方面起着至关重要的作用，并且可以作为长距离形态发生素发挥作用。理解 TGF-β/BMP 信号转导的动态和调控的关键是能够在生理表达水平和内源性时空模式下监测和操纵途径成分的工具。我们利用基因组工程技术，生成了一个全面的内源性表位或荧光标记受体、共受体、转录因子和 Drosophila BMP 和激活素信号通路关键反馈调节剂的文库。我们证明生成的等位基因具有生物活性，可用于评估相应蛋白质在组织和亚细胞水平的分布。此外，我们还表明，基因组平台可用于结构-功能和顺式调控分析。最后，我们提出了一组补充的基于蛋白质结合物的工具，这些工具可以可视化以及操纵表位标记蛋白的稳定性和亚细胞定位，为 BMP 信号转导及其他方面的分析提供了新的工具。
