Detection of Low-Abundance Metabolites in Live Cells Using an RNA Integrator
| Autor: | Jacob L. Litke, Samie R. Jaffrey, Mingxu You, Rigumula Wu |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
RNA Folding
Aptamer Clinical Biochemistry Biosensing Techniques Cleavage (embryo) 01 natural sciences Biochemistry Drug Discovery Escherichia coli RNA Catalytic Molecular Biology Fluorescent Dyes Pharmacology chemistry.chemical_classification biology Base Sequence 010405 organic chemistry Biomolecule Optical Imaging Ribozyme RNA Aptamers Nucleotide Small molecule 0104 chemical sciences Cell biology chemistry Integrator biology.protein Molecular Medicine Biosensor |
| Zdroj: | Cell chemical biology. 26(4) |
| ISSN: | 2451-9448 |
| Popis: | Summary Genetically encoded biosensors are useful tools for detecting the presence and levels of diverse biomolecules in living cells. However, low-abundance targets are difficult to detect because they are often unable to bind and activate enough biosensors to detect using standard microscopic imaging approaches. Here we describe a type of RNA-based biosensor, an RNA integrator, which enables detection of low-abundance targets in vitro and in living cells. The RNA integrator is an RNA sequence comprising a ribozyme and an unfolded form of the fluorogenic aptamer Broccoli. Upon binding its target, the ribozyme undergoes cleavage and releases Broccoli, which subsequently folds and becomes fluorescent. Importantly, each target molecule can bind and induce cleavage of multiple copies of the integrator sensor, resulting in an amplified signal. We show that this approach can be generalized to numerous different ribozyme types for the detection of various small molecules. |
| Databáze: | OpenAIRE |
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