Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia.
Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997 Moscow, Russia.
Int J Mol Sci. 2020 Dec 25;22(1):148. doi: 10.3390/ijms22010148.
Genetically-encoded fluorescent sensors have been actively developed over the last few decades and used in live imaging and drug screening. Real-time monitoring of drug action in a specific cellular compartment, organ, or tissue type; the ability to screen at the single-cell resolution; and the elimination of false-positive results caused by low drug bioavailability that is not detected by in vitro testing methods are a few of the obvious benefits of using genetically-encoded fluorescent sensors in drug screening. In combination with high-throughput screening (HTS), some genetically-encoded fluorescent sensors may provide high reproducibility and robustness to assays. We provide a brief overview of successful, perspective, and hopeful attempts at using genetically encoded fluorescent sensors in HTS of modulators of ion channels, Ca homeostasis, GPCR activity, and for screening cytotoxic, anticancer, and anti-parasitic compounds. We discuss the advantages of sensors in whole organism drug screening models and the perspectives of the combination of human disease modeling by CRISPR techniques with genetically encoded fluorescent sensors for drug screening.
在过去的几十年中，基因编码荧光传感器得到了积极的发展，并被用于活细胞成像和药物筛选。使用基因编码荧光传感器进行药物筛选具有明显的优势，例如可以实时监测特定细胞区室、器官或组织类型中的药物作用，以单细胞分辨率进行筛选，并消除由于体外测试方法无法检测到的低药物生物利用度而导致的假阳性结果。与高通量筛选（HTS）相结合，一些基因编码荧光传感器可能为测定提供高重现性和稳健性。我们简要概述了在离子通道调节剂、钙稳态、GPCR 活性的 HTS 中以及用于筛选细胞毒性、抗癌和抗寄生虫化合物中使用基因编码荧光传感器的成功、有前景和有希望的尝试。我们讨论了传感器在整个生物体药物筛选模型中的优势，以及通过 CRISPR 技术与基因编码荧光传感器相结合进行人类疾病建模用于药物筛选的前景。
