用于肌动蛋白成像的基于AsLOV2的光遗传学工具的设计与应用
Design and Use of AsLOV2-Based Optogenetic Tools for Actin Imaging.
作者信息
Kroll Kourtney L, Sosnick Tobin R, Rock Ronald S
机构信息
Department of Biochemistry and Molecular Biology & The Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL, USA.
出版信息
Methods Mol Biol. 2025;2840:89-100. doi: 10.1007/978-1-0716-4047-0_7.
We present protocols for using an optogenetic tool called LILAC for actin imaging. LILAC is a light-controlled version of Lifeact that uses the Avena sativa LOV2 (AsLOV2) domain. By significantly reducing Lifeact's affinity for the cytoskeleton in the dark, LILAC reduces concentration-dependent negative side effects while enabling new image processing methods. We discuss the considerations for using this probe of live-cell actin dynamics, including fluorescent protein selection, cell maintenance, microscopy protocols, and image processing. Our work highlights the potential of AsLOV2-based optogenetics for novel imaging and control tools in cell biology.
我们展示了使用一种名为LILAC的光遗传学工具进行肌动蛋白成像的实验方案。LILAC是Lifeact的光控版本，它使用了燕麦（Avena sativa）的LOV2（AsLOV2）结构域。通过在黑暗中显著降低Lifeact对细胞骨架的亲和力，LILAC减少了浓度依赖性负面效应，同时启用了新的图像处理方法。我们讨论了使用这种活细胞肌动蛋白动力学探针的注意事项，包括荧光蛋白选择、细胞维护、显微镜检查方案和图像处理。我们的工作突出了基于AsLOV2的光遗传学在细胞生物学新型成像和控制工具方面的潜力。
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