Center for Molecular Medicine, Molecular Cancer Research, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG Utrecht, the Netherlands.
STAR Protoc. 2021 Jan 13;2(1):100273. doi: 10.1016/j.xpro.2020.100273. eCollection 2021 Mar 19.
The relative positioning of organelles underlies fundamental cellular processes, including signaling, polarization, and cellular growth. Here, we describe the usage of a light-dependent heterodimerization system, LOVpep-ePDZ, to alter organelle positioning locally and reversibly in order to study the functional consequences of organelle positioning. The protocol gives details on how to accomplish expression of fusion proteins encoding this system, describes the imaging parameters to achieve subcellular activation in , and may be adapted for use in other model systems. For complete details on the use and execution of this protocol, please refer to De Henau et al. (2020).
细胞器的相对定位是基础细胞过程的基础，包括信号转导、极化和细胞生长。在这里，我们描述了使用光依赖性异二聚化系统 LOVpep-ePDZ 来局部和可逆地改变细胞器的定位，以研究细胞器定位的功能后果。该方案详细说明了如何完成编码该系统的融合蛋白的表达，描述了在 中实现亚细胞激活的成像参数，并可适用于其他模型系统。有关使用和执行此方案的完整详细信息，请参阅 De Henau 等人。（2020 年）。
