利用基因编码荧光生物传感器进行活体定量成像。

Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging.

机构信息

Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Nagoya, Japan.

Department of Biology, Stanford University, Stanford, CA, USA.

出版信息

Methods Mol Biol. 2021;2200:303-322. doi: 10.1007/978-1-0716-0880-7_14.

DOI:10.1007/978-1-0716-0880-7_14

PMID:33175384

Abstract

Fluorescent biosensors are powerful tools for tracking analytes or cellular processes in live organisms and allowing visualization of the spatial and temporal dynamics of cellular regulators. Fluorescent protein (FP)-based biosensors are extensively employed due to their high selectivity and low invasiveness. A variety of FP-based biosensors have been engineered and applied in plant research to visualize dynamic changes in pH, redox state, concentration of molecules (ions, sugars, peptides, ATP, reactive oxygen species, and phytohormones), and activity of transporters. In this chapter, we briefly summarize reported uses of FP-based biosensors in planta and show simple methods to monitor the dynamics of intracellular Ca in Arabidopsis thaliana using a ratiometric genetically encoded Ca indicator, MatryoshCaMP6s.

摘要

荧光生物传感器是追踪活体生物分析物或细胞过程的强大工具，并允许可视化细胞调节剂的时空动态。由于其高选择性和低侵入性，基于荧光蛋白 (FP) 的生物传感器得到了广泛的应用。已经设计和应用了各种基于 FP 的生物传感器来研究植物，以可视化 pH、氧化还原状态、分子浓度（离子、糖、肽、ATP、活性氧和植物激素）以及转运体活性的动态变化。在本章中，我们简要总结了报告中在植物中使用基于 FP 的生物传感器的情况，并展示了使用比率型遗传编码 Ca 指示剂 MatryoshCaMP6s 监测拟南芥细胞内 Ca 动态的简单方法。