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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Harmonizing the growing fluorogenic RNA aptamer toolbox for RNA detection and imaging.

Xiaocen Lu1, Kristen Y S Kong1, Peter J Unrau1

  • 1Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada. punrau@sfu.ca.

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|June 6, 2023
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Summary
This summary is machine-generated.

Fluorogenic RNA aptamers offer bright, tunable fluorescence for RNA detection and imaging, overcoming limitations of natural fluorescent molecules. This review details their selection, evaluation, and application in advanced biological imaging.

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Area of Science:

  • Biochemistry and Molecular Biology
  • Chemical Biology
  • Biotechnology

Background:

  • Naturally fluorescent RNA molecules are scarce, limiting RNA detection and imaging applications.
  • Fluorogenic RNA aptamers are engineered RNA molecules that exhibit enhanced fluorescence upon binding to specific ligands.
  • These aptamers offer comparable or superior molar brightness to fluorescent proteins, presenting a novel tool for biological research.

Purpose of the Study:

  • To review and discuss the selection methodologies employed for isolating novel fluorogenic RNA aptamers.
  • To provide a comprehensive evaluation of over seventy fluorogenic aptamer:ligand pairs based on key performance parameters.
  • To offer guidelines for selecting appropriate fluorescent RNA tools for specific applications, including single-molecule detection and multicolor imaging.

Main Methods:

  • Systematic review of literature on fluorogenic RNA aptamer selection and characterization.
  • Objective evaluation of identified aptamer:ligand pairs using parameters such as molar brightness, binding affinity, and fluorophore exchange.
  • Analysis of diverse fluorogenicity mechanisms and their implications for aptamer design and application.

Main Results:

  • Identification and evaluation of over seventy distinct fluorogenic RNA aptamer:ligand pairs.
  • Demonstration of various mechanisms driving fluorogenicity in RNA aptamer systems.
  • Compilation of objective data on aptamer performance metrics, facilitating comparative analysis.

Conclusions:

  • Fluorogenic RNA aptamers represent a powerful and versatile class of molecular tools for biological research.
  • Guidelines are provided for selecting optimal aptamers for applications like single-molecule detection and multicolor imaging.
  • The establishment of global standards for evaluating these systems is crucial for future development and standardization.