caa a22 4500 397575416 CHVBK 20180308164854.0 cr unu---uuuuu 161202e199612 xx s 000 0 eng 10.1093/hmg/5.12.1939 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.12.1939 Delivery of a Hammerhead Ribozyme Specifically Down-Regulates the Production of Fibrillin-1 by Cultured Dermal Fibroblasts [Elektronische Daten] [Michael W. Kilpatrick, Leonidas A. Phylactou, Maurice Godfrey, Catherine H. Wu, George Y. Wu, Petros Tsipouras] 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-1 gene (FBN1). Fibrillin is a 347 kDa glycoprotein that is a major constituent of the elastin-associated microfi-brils. 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 FBN1allele 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-RZ1 ribozyme into cultured dermal fibroblasts, by receptor-mediated endocytosis of a ribozyme-transferrin-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. © 1993 Oxford University Press Kilpatrick Michael W. Department of Pediatrics, University of Connecticut Health Center Farmington, CT 06030, USA aut Phylactou Leonidas A. Department of Pediatrics, University of Connecticut Health Center Farmington, CT 06030, USA aut Godfrey Maurice Department of Pediatrics and Munroe Center for Human Genetics, University of Nebraska Medical Center, Omaha, NE, USA aut Wu Catherine H. Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA aut Wu George Y. Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA aut Tsipouras Petros Department of Pediatrics, University of Connecticut Health Center Farmington, CT 06030, USA aut Human Molecular Genetics Oxford University Press 5/12(1996-12), 1939-1944 0964-6906 5:12<1939 1996 5 hmg https://doi.org/10.1093/hmg/5.12.1939 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.12.1939 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kilpatrick Michael W. Department of Pediatrics, University of Connecticut Health Center Farmington, CT 06030, USA aut NATIONALLICENCE 700 1- Phylactou Leonidas A. Department of Pediatrics, University of Connecticut Health Center Farmington, CT 06030, USA aut NATIONALLICENCE 700 1- Godfrey Maurice Department of Pediatrics and Munroe Center for Human Genetics, University of Nebraska Medical Center, Omaha, NE, USA aut NATIONALLICENCE 700 1- Wu Catherine H. Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA aut NATIONALLICENCE 700 1- Wu George Y. Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA aut NATIONALLICENCE 700 1- Tsipouras Petros Department of Pediatrics, University of Connecticut Health Center Farmington, CT 06030, USA aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/12(1996-12), 1939-1944 0964-6906 5:12<1939 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford