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Related Experiment Videos

Uridine insertion/deletion editing activities.

Jason Carnes1, Kenneth D Stuart

  • 1Seattle Biomedical Research Institute, Seattle, Washington, USA.

Methods in Enzymology
|July 31, 2007
PubMed
Summary
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This study details various assays for assessing RNA editing activity in trypanosomatids. Methods include in vitro assays for insertion/deletion editing and in vivo real-time PCR for monitoring editosome function.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Parasitology

Background:

  • RNA editing in trypanosomatids involves uridine insertion/deletion catalyzed by the editosome.
  • Assessing editosome activity is crucial for understanding RNA processing in these organisms.

Purpose of the Study:

  • To describe comprehensive assays for evaluating RNA editing and editosome activity.
  • To provide methods for both in vitro and in vivo assessments.

Main Methods:

  • In vitro assays: full-round and precleaved insertion/deletion editing, individual enzyme activities (TUTase, ligase, helicase).
  • In vivo assays: real-time PCR for simultaneous monitoring of multiple RNA targets.

Main Results:

  • Detailed protocols for various in vitro and in vivo RNA editing assays are presented.

Related Experiment Videos

  • Real-time PCR enables efficient, multi-target assessment of in vivo editing.

    • Conclusions:

    • The described assays provide robust tools for dissecting RNA editing mechanisms in trypanosomatids.
    • These methods facilitate the study of editosome function in both controlled laboratory settings and living organisms.