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Hammerhead Ribozyme-Mediated Knockdown of mRNA for Fibrotic Growth Factors: Transforming Growth Factor-Beta 1 and Connective Tissue Growth Factor

  • Paulette M. Robinson
  • Timothy D. Blalock
  • Rong Yuan
  • Alfred S. Lewin
  • Gregory S. SchultzEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 820)

Abstract

Excessive scarring (fibrosis) is a major cause of pathologies in multiple tissues, including lung, liver, kidney, heart, cornea, and skin. The transforming growth factor-β (TGF-β) system has been shown to play a key role in regulating the formation of scar tissue throughout the body. Furthermore, connective tissue growth factor (CTGF) has been shown to mediate most of the fibrotic actions of TGF-β, including stimulation of synthesis of extracellular matrix and differentiation of fibroblasts into myofibroblasts. Currently, no approved drugs selectively and specifically regulate scar formation. Thus, there is a need for a drug that selectively targets the TGF-β cascade at the molecular level and has minimal off-target side effects. This chapter focuses on the design of hammerhead ribozymes, measurement of kinetic activity, and assessment of knockdown mRNAs of TGF-β and CTGF in cell cultures.

Key words

Ribozymes TGF-β CTGF Scar formation Transduction Oligonucleotides 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Paulette M. Robinson
    • 1
  • Timothy D. Blalock
    • 2
  • Rong Yuan
    • 2
  • Alfred S. Lewin
    • 1
  • Gregory S. Schultz
    • 2
    Email author
  1. 1.Department of Molecular Genetics and MicrobiologyUniversity of FloridaGainesvilleUSA
  2. 2.Department of Obstetrics and Gynecology, College of Medicine, Institute for Wound ResearchUniversity of FloridaGainesvilleUSA

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