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Deoxyribozyme-catalyzed covalent labeling of RNAs

脱氧核酶催化的 RNA 共价标记

基本信息

批准号：
7306023
负责人：
DANA A BAUM
金额：
$ 4.67万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-28 至 2008-08-08
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Deoxyribozymes are DNA oligonucleotides that catalyze a variety of reactions. Through in vitro selection, deoxyribozymes have been identified that are able to join (ligate) two RNA substrates together via 2',5' linkages, forming branched RNA products. This catalytic activity opens the door to new methods of RNA derivatization and RNA labeling. The objective of this proposal is to develop deoxyribozymes that covalently link a derivatized RNA substrate (a "tagging" RNA) to target RNAs via 2',5'-branched linkages. These labeling deoxyribozymes will be identified by in vitro selection, with the goal of identifying deoxyribozymes that have minimal sequence requirements and are tolerant of useful biophysical labels such as fluorescein and biotin. These labeling deoxyribozymes are envisioned as general tools that could be used to label model RNAs for in vitro studies by methods such as FRET. Other uses are also possible by changing the functionality of the tagging RNA. Ultimately, these deoxyribozymes will be tested as mRNA labeling agents in vivo. The ability to covalently attach probe-containing tags to RNAs of interest in a cellular setting will be of great use for studies of development and disease.

描述（由申请人提供）：脱氧核酶是催化多种反应的DNA寡核苷酸。通过体外选择，脱氧核酶已被鉴定能够通过 2',5' 连接将两个 RNA 底物连接（连接）在一起，形成分支 RNA 产物。这种催化活性为 RNA 衍生化和 RNA 标记的新方法打开了大门。该提案的目的是开发脱氧核酶，通过 2',5'-分支连接将衍生化 RNA 底物（“标记”RNA）与靶 RNA 共价连接。这些标记脱氧核酶将通过体外选择进行鉴定，目的是鉴定具有最低序列要求并且能够耐受有用的生物物理标记（例如荧光素和生物素）的脱氧核酶。这些标记脱氧核酶被设想为通用工具，可用于通过 FRET 等方法标记模型 RNA，用于体外研究。通过改变标签 RNA 的功能也可以实现其他用途。最终，这些脱氧核酶将作为 mRNA 标记剂进行体内测试。在细胞环境中将含有探针的标签共价连接到感兴趣的 RNA 上的能力对于发育和疾病的研究将非常有用。