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Related Experiment Videos

Evolutionary dynamics and population control during in vitro selection and amplification with multiple targets.

Hua Shi1, Xiaochun Fan, Zhuoyu Ni

  • 1hs11@cornell.edu

RNA (New York, N.Y.)
|December 3, 2002
PubMed
Summary

Researchers tracked RNA aptamer evolution during selection experiments. They developed a method to control aptamer abundance, enabling exhaustive isolation of functional nucleic acid aptamers against multiple targets.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Nucleic Acid Aptamer Technology

Background:

  • In vitro selection and amplification are key for isolating functional nucleic acid aptamers from vast sequence libraries.
  • Simultaneous selection can yield aptamers against multiple targets, but controlling their relative abundance is challenging.

Purpose of the Study:

  • To analyze the evolutionary trajectories of two RNA aptamer families selected against distinct targets (protein and matrix).
  • To develop a method for controlling aptamer family abundance within selected pools.
  • To enable sequential and exhaustive isolation of aptamers against a mixture of targets.

Main Methods:

  • Iterative in vitro selection and amplification of RNA aptamers.
  • Tracking sequence abundance and binding activity across selection generations.

Related Experiment Videos

  • Employing specific ribonuclease digestion to modulate aptamer pool composition.

    • Main Results:

    • Two distinct RNA aptamer families against a protein (B52/SRp55) and a partitioning matrix were simultaneously isolated.
    • Aptamer families and individual members exhibited unique evolutionary paths during selection.
    • A ribonuclease digestion method successfully controlled the relative abundance of aptamer families.

    Conclusions:

    • The study elucidates the complex evolutionary dynamics during simultaneous aptamer selection.
    • A novel method using ribonuclease digestion offers control over aptamer abundance, reducing background selection.
    • This technique facilitates the comprehensive and sequential isolation of diverse aptamer populations from complex mixtures.