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Related Experiment Videos

Allosteric aptamers: targeted reversibly attenuated probes.

Xiangyu Cong1, Marit Nilsen-Hamilton

  • 1Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011, USA.

Biochemistry
|June 1, 2005
PubMed
Summary

A novel aptamer-based biosensor, the targeted reversibly attenuated probe (TRAP), utilizes structural flexibility to control aptamer-target binding. This TRAP system demonstrates sensitive, specific regulation for potential in vivo applications.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Biochemistry

Background:

  • Aptamers are nucleic acids with unique regulatory functions distinct from proteins.
  • Aptamers possess structural flexibility, enabling single-stranded folded structures for target binding and double-helical structures for sequence complementarity.

Purpose of the Study:

  • To develop an aptamer-based biosensor (TRAP) leveraging aptamer structural flexibility.
  • To investigate the regulatory mechanism of a cis-complementary nucleic acid (attenuator) on aptamer-target interaction.
  • To explore the potential of TRAP for in vivo applications.

Main Methods:

  • Development of a targeted reversibly attenuated probe (TRAP) integrating an aptamer and an attenuator.
  • Utilizing a target nucleic acid (regNA) to hybridize with the TRAP's central sequence, thereby regulating aptamer activity.

Related Experiment Videos

  • Assessing TRAP activation sensitivity to single-base mismatches at physiological temperatures.

    • Main Results:

    • The attenuator not only inhibits aptamer binding but also acts as a chaperone, promoting TRAP folding for regNA activation.
    • TRAP activation by complementary nucleic acid is highly sensitive to single-base mismatches at physiological temperatures.
    • The ATP-DNA TRAP system demonstrated effective regulation of aptamer activity.

    Conclusions:

    • TRAP biosensors offer a novel mechanism for regulating aptamer-target interactions through nucleic acid hybridization.
    • The chaperone-like function of the attenuator enhances TRAP efficiency and specificity.
    • TRAP technology holds promise for in vivo applications such as enzyme regulation, pathway modulation, and gene expression imaging.