In vitro selection methodologies to probe RNA function and structure

被引:33
作者
Conrad, RC [1 ]
Baskerville, S [1 ]
Ellington, AD [1 ]
机构
[1] INDIANA UNIV,DEPT CHEM,BLOOMINGTON,IN 47405
关键词
aptamer; in vitro selection; SELEX; Molecular evolution; HTLV-I; rex; rex-binding element;
D O I
10.1007/BF01715810
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
In vitro selection, or SELEX, has been used both to characterize the interaction of natural nucleic acids with proteins and to generate novel nucleic acid-binding species, or aptamers. Although numerous reports have demonstrated the power of the technique, they have not expanded on the methodologies that can be used for selection. This review focuses on the considerations and problems involved in selecting protein-binding aptamers from a random-sequence RNA pool. As an illustration, we describe two approaches to selecting aptamers to a particular target, the HTLV-I Rex protein. In the first, complete randomization is used to find an artificial, high-affinity RNA binding site. In the second, the contributions of individual nucleotides and ! or base pairs to the natural Rex-binding element are determined by mutating the wild-type sequence and selecting active binding variants.
引用
收藏
页码:69 / 78
页数:10
相关论文
共 50 条
[1]  
ABMAYR SB, 1988, CURRENT PROTOCOLS MO, V2
[2]   STRUCTURE-FUNCTION ANALYSES OF THE HTLV-I REX AND HIV-1 REV RNA RESPONSE ELEMENTS - INSIGHTS INTO THE MECHANISM OF REX AND REV ACTION [J].
AHMED, YF ;
HANLY, SM ;
MALIM, MH ;
CULLEN, BR ;
GREENE, WC .
GENES & DEVELOPMENT, 1990, 4 (06) :1014-1022
[3]   2'-FLUORO-2'-DEOXYNUCLEOSIDE AND 2'-AMINO-2'-DEOXYNUCLEOSIDE 5'-TRIPHOSPHATES AS SUBSTRATES FOR T7 RNA-POLYMERASE [J].
AURUP, H ;
WILLIAMS, DM ;
ECKSTEIN, F .
BIOCHEMISTRY, 1992, 31 (40) :9636-9641
[4]   HIV-1 REV REGULATION INVOLVES RECOGNITION OF NON-WATSON-CRICK BASE-PAIRS IN VIRAL-RNA [J].
BARTEL, DP ;
ZAPP, ML ;
GREEN, MR ;
SZOSTAK, JW .
CELL, 1991, 67 (03) :529-536
[5]  
BASKERVILLE S, 1995, IN PRESS J VIROLOGY
[6]   SELECTION OF SINGLE-STRANDED-DNA MOLECULES THAT BIND AND INHIBIT HUMAN THROMBIN [J].
BOCK, LC ;
GRIFFIN, LC ;
LATHAM, JA ;
VERMAAS, EH ;
TOOLE, JJ .
NATURE, 1992, 355 (6360) :564-566
[7]   THE TYPE-I HUMAN T-CELL LEUKEMIA-VIRUS (HTLV-I) REX TRANSACTIVATOR BINDS DIRECTLY TO THE HTLV-I REX AND THE TYPE-1 HUMAN-IMMUNODEFICIENCY-VIRUS REV RNA RESPONSE ELEMENTS [J].
BOGERD, HP ;
HUCKABY, GL ;
AHMED, YF ;
HANLY, SM ;
GREENE, WC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (13) :5704-5708
[8]   SELECTION OF HIGH-AFFINITY RNA LIGANDS TO REVERSE-TRANSCRIPTASE - INHIBITION OF CDNA SYNTHESIS AND RNASE-H ACTIVITY [J].
CHEN, H ;
GOLD, L .
BIOCHEMISTRY, 1994, 33 (29) :8746-8756
[9]  
CONRAD R, 1994, J BIOL CHEM, V269, P32051
[10]  
CONRAD RC, 1995, IN PRESS METHODS ENZ