Intracellular ribozyme applications
Intracellular ribozyme applications
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DOI：
10.1042/bst0301140
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发表时间：
2002-11-01
影响因子：
3.9
通讯作者：
Rossi, JJ
中科院分区：
文献类型：
--
作者：
Castanotto, D;Li, JR;Rossi, JJ
反式作用核酶的精细靶标选择性促进了它们作为潜在的治疗剂和下调细胞转录物的工具的使用。在活细胞中，rna的自由扩散是极其有限的，如果它存在的话。因此，使核酶与其同源靶标碱基对需要将核酶转录物与靶RNA共定位。此外，并不是一个给定的目标RNA上的所有位点都可以被核酶碱基配对。细胞蛋白极大地影响RNA的运输和结构，因此使核酶在细胞中有效工作是一个重大挑战。本文讨论了如何使工程核酶有效地与细胞中的靶标配对和切割。这里描述的工作阐明了天然mrna的靶位点选择方法，核酶表达方法，以及获得核酶的离散胞内定位的策略。
The exquisite target selectivity of trans-acting ribozymes has fostered their use as potential therapeutic agents and tools for down-regulating cellular transcripts. In living cells, free diffusion of RNAs is extremely limited, if it exists at all. Thus, getting ribozymes to base-pair with their cognate targets requires co-localizing the ribozyme transcript with the target RNA. In addition, not all sites along a given target RNA are equally accessible to ribozyme base pairing. Cellular proteins greatly influence the trafficking and structure of RNA, and therefore making ribozymes work effectively in cells a significant challenge. This article addresses the problems of getting engineered ribozymes to effectively pair with and cleave targets in cells. The work described here illuminates methods for target-site selection on native mRNAs, methods for ribozyme expression, and strategies for obtaining a discrete intracellular localization of ribozymes.