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A circular trans-acting hepatitis delta virus ribozyme

M Puttaraju¹, A T Perrotta, M D Been

¹Department of Biochemistry, Duke University Medical Center, Durham, NC 27710.

Nucleic Acids Research

|September 11, 1993

Summary

Circular ribozymes, based on the hepatitis delta virus (HDV) motif, show comparable activity and enhanced stability against nucleases. Circularization offers a promising strategy for stabilizing ribozymes.

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Area of Science:

Molecular Biology
Biochemistry
RNA Therapeutics

Background:

Ribozymes are catalytic RNA molecules with therapeutic potential.
Hepatitis delta virus (HDV) ribozymes are well-characterized for their catalytic activity.
Stabilizing ribozymes against degradation is crucial for their application.

Purpose of the Study:

To produce and characterize a circular trans-acting ribozyme based on the HDV motif.
To assess the catalytic activity and nuclease resistance of the circular ribozyme.
To evaluate circularization as a stabilization strategy for ribozymes.

Main Methods:

In vitro production of a circular ribozyme via splicing of a modified group I intron precursor.
Assay of trans-cleavage activity of the circular ribozyme and linear controls.
Assessment of nuclease resistance in nuclear and cytoplasmic HeLa cell extracts.

Main Results:

A circular HDV-like ribozyme was successfully produced.
Trans-cleavage activity of the circular ribozyme was comparable to linear variants.
The circular ribozyme exhibited enhanced resistance to nuclease degradation compared to its linear counterpart.

Conclusions:

Circularization of HDV-like ribozymes maintains catalytic activity.
Circularization enhances ribozyme stability against cellular nucleases.
This suggests circularization is a viable alternative to chemical modification for ribozyme stabilization.

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