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Related Concept Videos

Ribozymes02:47

Ribozymes

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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can...
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Ribozymes02:47

Ribozymes

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Engineering a ribozyme cleavage-induced split fluorescent aptamer complementation assay.

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|March 5, 2016
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Summary
This summary is machine-generated.

Researchers developed a new assay to measure hammerhead ribozyme activity in real-time. This technology validates RNA loop-loop interactions, crucial for engineering gene-regulating genetic switches.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Therapeutics

Background:

  • Hammerhead ribozymes are RNA enzymes that self-cleave, regulating gene expression.
  • Their function depends on loop-loop interactions, which are difficult to predict.
  • Accurate measurement of ribozyme activity is vital for genetic switch engineering.

Purpose of the Study:

  • To develop a real-time assay for measuring hammerhead ribozyme self-cleavage activity.
  • To enable the validation of RNA loop-loop interactions in hammerhead ribozymes.
  • To facilitate the rational design of novel ribozyme-based genetic switches.

Main Methods:

  • Engineered RNA architectures incorporating a split Spinach aptamer.
  • Real-time measurement of ribozyme self-cleavage activity in vitro.
  • Strand displacement mechanism to complement the Spinach aptamer upon cleavage.
  • Correlation of ribozyme cleavage with Spinach fluorescence.

Main Results:

  • Developed a novel, fully RNA-based assay for hammerhead ribozyme performance.
  • Demonstrated real-time measurement of self-cleavage activity.
  • Established a correlation between ribozyme cleavage and Spinach fluorescence.
  • Provided a straightforward method for validating loop-loop interactions.

Conclusions:

  • The developed assay offers a rapid and efficient technology for assessing hammerhead ribozyme function.
  • This method is crucial for understanding and engineering RNA-based genetic switches.
  • The assay facilitates the validation of critical loop-loop interactions in ribozyme design.