生物传感器
适体
氟化物
核糖开关
核糖核酸
化学
试剂
纳米技术
荧光
组合化学
生物化学
材料科学
生物
分子生物学
无机化学
非编码RNA
有机化学
物理
基因
量子力学
作者
Radi Khodr,Claire Husser,Michaël Ryckelynck
出处
期刊:Methods in Enzymology
日期:2024-01-01
卷期号:: 85-107
标识
DOI:10.1016/bs.mie.2023.12.019
摘要
Fluorinated compounds, whether naturally occurring or from anthropogenic origin, have been extensively exploited in the last century. Degradation of these compounds by physical or biochemical processes is expected to result in the release of fluoride. Several fluoride detection mechanisms have been previously described. However, most of these methods are not compatible with high- and ultrahigh-throughput screening technologies, lack the ability to real-time monitor the increase of fluoride concentration in solution, or rely on costly reagents (such as cell-free expression systems). Our group recently developed "FluorMango" as the first completely RNA-based and direct fluoride-specific fluorogenic biosensor. To do so, we merged and engineered the Mango-III light-up RNA aptamer and the fluoride-specific aptamer derived from a riboswitch, crcB. In this chapter, we explain how this RNA-based biosensor can be produced in large scale before providing examples of how it can be used to quantitatively detect (end-point measurement) or monitor in real-time fluoride release in complex biological systems by translating it into measurable fluorescent signal.
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