RNA complex purification using high-affinity fluorescent RNA aptamer tags

被引:12
|
作者
Panchapakesan, Shanker Shyam S. [1 ]
Jeng, Sunny C. Y. [1 ]
Unrau, Peter J. [1 ]
机构
[1] Simon Fraser Univ, Dept Mol Biol & Biochem, Burnaby, BC V5A 1S6, Canada
来源
关键词
RNA aptamers; RNA purification tags; in vitro selection; RNA Mango; binding affinity; fluorescent enhancement; MESSENGER-RNA; BINDING PROTEIN; IDENTIFICATION; EXPRESSION; PARTICLES; MIMICS;
D O I
10.1111/nyas.12663
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
RNA plays important roles in cellular processes, but RNA-protein complexes are notoriously hard to isolate and study. We compare and contrast existing RNA- and protein-purification strategies with the potential of new RNA-tagging systems such as RNA Spinach and RNA Mango. Each RNA aptamer binds a small fluorophore, resulting in a highly fluorescent complex that is thousands of times brighter than the unbound fluorophore. Provided that the aptamer binding affinity is high enough, derivatized dyes can be used in conjunction with these aptamers to purify RNA complexes while simultaneously using their intrinsic fluorescence to track the complex of interest. The known strengths and weakness of these RNA tagging systems are discussed.
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页码:149 / 155
页数:7
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