锤头状核酶介导的纤维化生长因子mRNA敲低：转化生长因子-β1和结缔组织生长因子。
Hammerhead ribozyme-mediated knockdown of mRNA for fibrotic growth factors: transforming growth factor-beta 1 and connective tissue growth factor.
作者信息
Robinson Paulette M, Blalock Timothy D, Yuan Rong, Lewin Alfred S, Schultz Gregory S
机构信息
Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL, USA.
出版信息
Methods Mol Biol. 2012;820:117-32. doi: 10.1007/978-1-61779-439-1_8.
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.
过度瘢痕形成（纤维化）是包括肺、肝、肾、心、角膜和皮肤在内的多种组织发生病变的主要原因。转化生长因子-β（TGF-β）系统已被证明在调节全身瘢痕组织形成中起关键作用。此外，结缔组织生长因子（CTGF）已被证明介导TGF-β的大多数纤维化作用，包括刺激细胞外基质合成和成纤维细胞分化为肌成纤维细胞。目前，尚无获批的药物能选择性且特异性地调节瘢痕形成。因此，需要一种在分子水平上选择性靶向TGF-β级联反应且脱靶副作用最小的药物。本章重点介绍锤头状核酶的设计、动力学活性的测定以及细胞培养中TGF-β和CTGF的mRNA敲低评估。
Zotero 插件