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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

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Related Experiment Video

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Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
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Programming ADAR-recruiting hairpin RNA sensor to detect endogenous molecules.

Pei-Pei Qin1, Pin-Ru Chen1, Liu Tan1

  • 1Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, No.18 Chao Wang Road, Gongshu District, Hangzhou 310014, China.

Nucleic Acids Research
|December 14, 2024
PubMed
Summary

Researchers developed a versatile hairpin RNA sensor for detecting multiple intracellular molecules. This novel ADAR-based biosensor offers enhanced sensitivity and specificity for RNA, small molecules, and proteins in live cells.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biosensing

Background:

  • Adenosine deaminases acting on RNA (ADARs) show potential for in vivo biosensing.
  • Current ADAR sensors are limited to single-target detection, restricting their application scope.

Purpose of the Study:

  • To develop a novel hairpin RNA sensor for multiplexed in vivo detection.
  • To overcome limitations of single-target ADAR sensors.
  • To expand the range of detectable intracellular molecules.

Main Methods:

  • Engineered hairpin RNA structures to create intramolecular duplexes for ADAR recruitment and editing.
  • Utilized base-pairing interactions for enhanced stability and reverse effects for sensitive detection.
  • Integrated RNA aptamers for detecting ATP and NF-κB via conformational changes.

Main Results:

  • Demonstrated detection of messenger RNA, single-nucleotide variants, ATP, and NF-κB in live cells.
  • Achieved high sensitivity and specificity irrespective of sequence requirements.
  • Showcased the sensor's adaptability for diverse intracellular molecule detection.

Conclusions:

  • The developed hairpin RNA sensor broadens the scope of ADAR-based biosensing.
  • This technology enables multiplexed detection and offers potential for cell manipulation.
  • Represents a significant advancement in in vivo biosensing and molecular detection.