Delivery of a hammerhead ribozyme specifically down-regulates the production of fibrillin-1 by cultured dermal fibroblasts

Michael W. Kilpatrick, Leonidas A. Phylactou, Maurice Godfrey, Catherine H. Wu, George Y. Wu, Petros Tsipouras

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The hammerhead ribozyme is a small catalytic RNA molecule. Potential hammerhead ribozymes that possess a catalytic domain and flanking sequence complementary to a target mRNA can cleave in trans at a putative cleavage site within the target molecule. We have investigated the potential of hammerhead ribozymes to down-regulate the product of the fibrillin-l gene (FBN1). Fibrillin is a 347 kDa glycoprotein that is a major constituent of the elastin-associated microfibrils. Mutations in the FBN1 gene are responsible for Marfan syndrome (MFS), a common systemic disorder of the connective tissue. Many FBN1 mutations responsible for MFS appear to act in a dominant-negative fashion, raising the possibility that reduction of the amount of product from the mutant FBN1 allele might be a valid therapeutic approach for MFS. A trans-acting hammerhead ribozyme (FBN1-RZ1) targeted to the 5' end of the human FBN1 mRNA has been designed and synthesized, and shown to cleave its target efficiently in vitro. FBN1-RZ1 cleavage is magnesium dependent and efficient at both 37 and 50°C. Delivery of the FBN1-RZI ribozyme into cultured dermal fibroblasts, by receptor-mediated endocytosis of a ribozymetransferrin-polylysine complex, specifically reduces both cellular FBN1 mRNA and the deposition of fibrillin in the extracellular matrix. These results suggest that the use of hammerhead ribozymes is a valid approach to the study of fibrillin gene expression and possibly to the development of a therapeutic approach to MFS.

Original languageEnglish (US)
Pages (from-to)1939-1944
Number of pages6
JournalHuman Molecular Genetics
Volume5
Issue number12
StatePublished - Dec 1 1996

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Marfan Syndrome
Down-Regulation
Fibroblasts
Skin
Catalytic RNA
Messenger RNA
Microfibrils
Mutation
Polylysine
Elastin
Endocytosis
Connective Tissue
Magnesium
Genes
Extracellular Matrix
Catalytic Domain
Glycoproteins
Alleles
hammerhead ribozyme
Fibrillin-1

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

Cite this

Delivery of a hammerhead ribozyme specifically down-regulates the production of fibrillin-1 by cultured dermal fibroblasts. / Kilpatrick, Michael W.; Phylactou, Leonidas A.; Godfrey, Maurice; Wu, Catherine H.; Wu, George Y.; Tsipouras, Petros.

In: Human Molecular Genetics, Vol. 5, No. 12, 01.12.1996, p. 1939-1944.

Research output: Contribution to journalArticle

Kilpatrick, Michael W. ; Phylactou, Leonidas A. ; Godfrey, Maurice ; Wu, Catherine H. ; Wu, George Y. ; Tsipouras, Petros. / Delivery of a hammerhead ribozyme specifically down-regulates the production of fibrillin-1 by cultured dermal fibroblasts. In: Human Molecular Genetics. 1996 ; Vol. 5, No. 12. pp. 1939-1944.
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