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.
引用
下载
收藏
页码:149 / 155
页数:7
相关论文
共 50 条
  • [41] SnAvi - a new tandem tag for high-affinity protein-complex purification
    Schaeffer, Ursula
    Schlosser, Andreas
    Mueller, Kristian M.
    Schaefer, Angelika
    Katava, Nenad
    Baumeister, Ralf
    Schulze, Ekkehard
    NUCLEIC ACIDS RESEARCH, 2010, 38 (06) : e91.1 - e91.13
  • [42] LNA (Locked nucleic acid): High-affinity targeting of complementary RNA and DNA
    Vester, B
    Wengel, J
    BIOCHEMISTRY, 2004, 43 (42) : 13233 - 13241
  • [43] BINDING AND STRUCTURAL-ANALYSIS OF HIGH-AFFINITY RNA LIGANDS TO HUMAN THROMBIN
    MCBRIDE, B
    SCHNEIDER, D
    DAKIS, K
    TASSET, D
    BLOOD, 1993, 82 (10) : A276 - A276
  • [44] Isolation of high-affinity GTP aptamers from partially structured RNA libraries
    Davis, JH
    Szostak, JW
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (18) : 11616 - 11621
  • [45] Structure of an RNA aptamer in complex with the fluorophore tetramethylrhodamine
    Duchardt-Ferner, Elke
    Juen, Michael
    Bourgeois, Benjamin
    Madl, Tobias
    Kreutz, Christoph
    Ohlenschlaeger, Oliver
    Woehnert, Jens
    NUCLEIC ACIDS RESEARCH, 2020, 48 (02) : 949 - 961
  • [46] Solution structure of the tobramycin–RNA aptamer complex
    Licong Jiang
    Dinshaw J. Patel
    Nature Structural Biology, 1998, 5 : 769 - 774
  • [47] AFFINITY PURIFICATION OF AMINOACYL-TRANSFER RNA
    LOUIE, A
    MASUDA, E
    YODER, M
    JURNAK, F
    ANALYTICAL BIOCHEMISTRY, 1984, 141 (02) : 402 - 408
  • [48] PURIFICATION AND RECONSTITUTION OF THE HIGH-AFFINITY CHOLINE TRANSPORTER
    KNIPPER, M
    KAHLE, C
    BREER, H
    BIOCHIMICA ET BIOPHYSICA ACTA, 1991, 1065 (02) : 107 - 113
  • [49] Engineering high-affinity scaffolds for protein purification
    Liu, Fang
    Chen, Wilfred
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2012, 243
  • [50] Recruiting the Immune System against Pathogenic Bacteria Using High-Affinity Chimeric Tags
    Belo, Yael
    Malach, Einav
    Hayouka, Zvi
    Bioconjugate Chemistry, 2024, 35 (11) : 1716 - 1722