通过使用2'，3'-环磷酸RNA底物的锌离子依赖性脱氧核酶控制RNA连接中位点选择性和区域选择性的方向。
Controlling the direction of site-selectivity and regioselectivity in RNA ligation by Zn2+-dependent deoxyribozymes that use 2',3'-cyclic phosphate RNA substrates.
作者信息
Kost Diana M, Gerdt Joseph P, Pradeepkumar P I, Silverman Scott K
机构信息
Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, USA.
出版信息
Org Biomol Chem. 2008 Dec 7;6(23):4391-8. doi: 10.1039/b813566e. Epub 2008 Oct 27.
Abstract
Our previous efforts have used in vitro selection to identify numerous Zn(2+)-dependent deoxyribozymes that ligate two RNA substrates with reaction at a 2',3'-cyclic phosphate. Each deoxyribozyme creates one of several different RNA linkages, including native 3'-5' and non-native 2'-5' phosphodiester bonds as well as many unnatural linkages. In this report, we describe experiments to reveal design aspects of the selection strategy that favor site-selective and regioselective synthesis of native 3'-5' RNA linkages. The results also reveal that an explicit selection pressure for RNA substrate sequence generality must be developed if the deoxyribozymes are to have practical generality.
摘要
我们之前的工作利用体外筛选来鉴定众多依赖锌离子的脱氧核酶，这些酶能在2',3'-环磷酸处连接两个RNA底物并发生反应。每个脱氧核酶会形成几种不同RNA连接方式中的一种，包括天然的3'-5'磷酸二酯键、非天然的2'-5'磷酸二酯键以及许多非天然连接方式。在本报告中，我们描述了相关实验，以揭示有利于天然3'-5' RNA连接位点选择性和区域选择性合成的筛选策略设计方面。结果还表明，如果脱氧核酶要具有实际通用性，就必须对RNA底物序列通用性施加明确的筛选压力。
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