Lipofectin‐aided cell delivery of ribozyme targeted to human urokinase receptor mRNA

FEBS Letters

Megyeri, Katalin¹  Karikó, Katalin¹  Xiao, Qi¹  Barnathan, Elliot S.¹ 

[1] Cardiovascular Division, Department of Medicine, University of Pennsylvania School of Medicine, 3610 Hamilton Walk, Philadelphia, PA 19104-6060, USA

关键词: Hammerhead ribozyme;    Delivery;    Stability;    Urokinase receptor;    Lipofectin;    HOS cell;   

DOI  :  10.1016/0014-5793(94)00914-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A 37-mer hammerhead ribozyme has been designed to efficiently cleave the 1.4 kb mRNA of the urokinase plasminogen activator receptor (uPAR). Under in vitro conditions, the chemically synthesized ribozyme cleaved uPAR mRNA and inhibited its translation in a concentration-dependent fashion. The ribozymes were 5′-[³⁵S]thiophosphorylated and used as a model to analyze conditions for RNA delivery in a cultured human osteosarcoma cell system. Ribozymes degraded immediately in cell-conditioned medium but ribozymes complexed with lipofectin were protected from RNases for up to 22 h. Lipofectin rapidly transported ribozyme into the cell, where it accumulated almost exclusively in the cytoplasm. Thus, lipofectin dramatically enhances stability and cytoplasmic delivery of ribozymes, potentially enabling targeting of mRNA in vivo.

【 授权许可】

Unknown