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J. Biochem, 2004, Vol. 136, No. 2 133-147
© 2004 The Japanese Biochemical Society

JB REVIEWS

Ribozymes: Applications to Functional Analysis and Gene Discovery

Maki Shiota1,2, Masayuki Sano2, Makoto Miyagishi1,2 and Kazunari Taira1,2,*

1 Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Tokyo 113-8656; and 2 Gene Function Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba 305-8562

ABSTRACT

Ribozymes are catalytic RNA molecules that cleave RNAs with high specificity. Since the discovery of these non-protein enzymes, the rapidly developing field of ribozymes has been of particular interest because of the potential utility of ribozymes as tools for reversed genetics. However, despite extensive efforts, the activity of ribozymes in vivo has not usually been high enough to achieve the desirable biological effects. Now, by the use of RNA polymerase III (pol III) promoters, the ribozyme activity in cells has been successfully improved by developing efficient transport systems for the transcripts to the cytoplasm. In addition, it is possible to cleave a specific target RNA in cells by using an allosterically controllable ribozyme or an RNA-protein hybrid ribozyme. These ribozymes are potentially applicable to molecular gene therapy and efficient gene discovery systems. Furthermore, the developed pol III expression system is applicable to the expression of small interfering RNAs (siRNAs). The advantage of such ribozymes over siRNAs is the high specificity of the ribozyme that would not cause interferon responses.

FOOTNOTES

* To whom correspondence should be addressed. Tel: +81-3-5841-8828 or +81-29-861-3015, Fax: +81-3-5841-7340 or +81-29-861-3019, E-mail: taira{at}chembio.t.u-tokyo.ac.jp


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