RNA Group/Groupe ARN, Département de Biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke , Sherbrooke, Québec, Canada.
Nucleic Acid Ther. 2011 Aug;21(4):241-52. doi: 10.1089/nat.2011.0301. Epub 2011 Jul 8.
The Hepatitis Delta Virus (HDV) ribozyme, which is well adapted to the environment of the human cell, is an excellent candidate for the future development of gene-inactivation systems. On top of this, a new generation of HDV ribozymes now exists that benefits from the addition of a specific on/off adaptor (specifically the SOFA-HDV ribozymes) which greatly increases both the ribozyme's specificity and its cleavage activity. Unlike RNAi and hammerhead ribozymes, the designing of SOFA-HDV ribozymes to cleave, in trans, given RNA species has never been the object of a systematic optimization study, even with their recent use for the gene knockdown of various targets. This report aims at both improving and clarifying the design process of SOFA-HDV ribozymes. Both the ribozyme and the targeted RNA substrate were analyzed in order to provide new criteria that are useful in the selection of the most potent SOFA-HDV ribozymes. The crucial features present in both the ribozyme's biosensor and blocker, as well as at the target site, were identified and characterized. Simple rules were derived and tested using hepatitis C virus NS5B RNA as a model target. Overall, this method should promote the use of the SOFA-HDV ribozymes in a plethora of applications in both functional genomics and gene therapy.
乙型肝炎 Delta 病毒（HDV）核酶非常适应人体细胞的环境，是未来基因失活系统发展的理想候选者。此外，现在已经出现了新一代的 HDV 核酶，它们受益于添加了一种特定的开/关适应体（具体为 SOFA-HDV 核酶），这大大提高了核酶的特异性和切割活性。与 RNAi 和锤头核酶不同，针对特定 RNA 种类进行切割的 SOFA-HDV 核酶的设计从未成为系统优化研究的对象，尽管它们最近被用于各种靶标的基因敲低。本报告旨在改进和阐明 SOFA-HDV 核酶的设计过程。对核酶和靶向 RNA 底物进行了分析，以提供有用的新标准，用于选择最有效的 SOFA-HDV 核酶。确定并描述了核酶的生物传感器和阻断剂以及靶位点中存在的关键特征。使用丙型肝炎病毒 NS5B RNA 作为模型靶标，推导出了简单的规则并进行了测试。总的来说，这种方法应该促进 SOFA-HDV 核酶在功能基因组学和基因治疗中的大量应用。
