Buckley Clare E
Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
Methods Mol Biol. 2019;1920:143-162. doi: 10.1007/978-1-4939-9009-2_10.
This chapter describes the use of optogenetic heterodimerization in single cells within whole-vertebrate embryos. This method allows the use of light to reversibly bind together an "anchor" protein and a "bait" protein. Proteins can therefore be directed to specific subcellular compartments, altering biological processes such as cell polarity and signaling. I detail methods for achieving transient expression of fusion proteins encoding the phytochrome heterodimerization system in early zebrafish embryos (Buckley et al., Dev Cell 36(1):117-126, 2016) and describe the imaging parameters used to achieve subcellular light patterning.
本章描述了光遗传学异源二聚化在全脊椎动物胚胎单细胞中的应用。该方法允许利用光将“锚定”蛋白和“诱饵”蛋白可逆地结合在一起。因此，蛋白质可以被引导至特定的亚细胞区室，从而改变诸如细胞极性和信号传导等生物学过程。我详细介绍了在斑马鱼早期胚胎中实现编码光敏色素异源二聚化系统的融合蛋白瞬时表达的方法（Buckley等人，《发育细胞》36(1):117 - 126，2016年），并描述了用于实现亚细胞光图案化的成像参数。
