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Delivery of unmodified bioactive ribozymes by an RNA-stabilizing polyethylenimine (LMW-PEI) efficiently down-regulates gene expression

Gene Therapy volume 9, pages 1700–1707 (2002)Cite this article

Abstract

The sequence-specific cleavage of RNA molecules through ribozyme targeting is particularly attractive since it allows the effective abrogation of protein expression. So far, however, use of enzymatically active RNA molecules (ribozymes) has, without chemical modification, been severely hampered by ribozyme instability and poor cellular uptake. In this paper, we present a method for protection and cellular delivery of ribozymes by complexation with a low molecular weight polyethylenimine (LMW-PEI). We show that LMW-PEI almost completely stabilizes ribozymes or any RNA against degradation in vitro. Upon their highly efficient cellular uptake, non-toxic LMW-PEI-complexed ribozymes display intracellular bioactivity already at low concentrations as demonstrated by down-regulation of two different genes in different cell lines. In vivo, LMW-PEI-complexed ribozymes were stabilized after intraperitoneal (i.p.) injections, showed prolonged circulation time and intact ribozymes were detected in the subcutaneous (s.c.) tumor mass 60 min after the injection. In addition, i.p. injections of LMW-PEI-complexed ribozymes targeted against the growth factor pleiotrophin (PTN) resulted in marked reduction of s.c. human melanoma tumor growth and of intratumoral PTN levels in a mouse xenograft model. Thus, this paper describes a novel method for exogenous delivery of any bioactive RNA ribozyme in vitro and in vivo without chemical modification.

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Acknowledgements

The authors are grateful to Prof. R Moll, Chairman of the department of Pathology, Philipps-University Marburg, for expert microscopic evaluation of the tumor sections. The authors would like to thank Andrea Wüstenhagen, Daniela Wagner, Helga Radler and Barbara Siegel for expert technical assistance. This work was supported by a grant from the Philipps-University Forschungspool to AA.

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  1. A Aigner and D Fischer: The first two authors contributed equally to the work

Authors and Affiliations

  1. Department of Pharmacology and Toxicology, Philipps-University Marburg, Germany

    A Aigner & F Czubayko

  2. Department of Pharmaceutical Technology and Biopharmacy, Philipps-University Marburg, Germany

    D Fischer, T Merdan, C Brus & T Kissel

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  1. A Aigner
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  2. D Fischer
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  3. T Merdan
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Aigner, A., Fischer, D., Merdan, T. et al. Delivery of unmodified bioactive ribozymes by an RNA-stabilizing polyethylenimine (LMW-PEI) efficiently down-regulates gene expression. Gene Ther 9, 1700–1707 (2002). https://doi.org/10.1038/sj.gt.3301839

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