Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs

被引：48

作者：

Moriyama, K

Kimoto, M

Mitsui, T

Yokoyama, S

Hirao, I

机构：

[1] Univ Tokyo, Adv Sci & Technol Res Ctr, Meguro Ku, Tokyo 1538904, Japan

[2] RIKEN, Genom Sci Ctr, Prot Res Grp, Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan

[3] Univ Tokyo, Dept Biophys & Biochem, Grad Sch Sci, Bunkyo Ku, Tokyo 1130033, Japan

[4] RIKEN, Harima Inst, SPring 8, Sayo, Hyogo 6795148, Japan

来源：

NUCLEIC ACIDS RESEARCH | 2005年 / 33卷 / 15期

关键词：

D O I：

10.1093/nar/gni128

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：

Direct site-specific biotinylation of RNA molecules was achieved by specific transcription mediated by unnatural base pairs. Unnatural base pairs between 2-amino-6-(2-thienyl)purine (denoted by s) and 2-oxo(1H)pyridine (denoted by y), or 2-amino-6-(2-thiazolyl)purine (denoted as v) and y specifically function in T7 transcription. Using these unnatural base pairs, the substrate of biotinylated-y (Bio-yTP) was selectively incorporated into RNA, opposite s or v in the DNA templates, by T7 RNA polymerase. This method was applied to the immobilization of an RNA aptamer on sensor chips, and the aptamer accurately recognized its target protein. This direct site-specific biotinylation will provide a tool for RNA-based biotechnologies.

引用

收藏

页码：1 / 8

页数：8

相关论文

共 16 条

[1] BIOCYTIN HYDRAZIDE - A SELECTIVE LABEL FOR SIALIC ACIDS, GALACTOSE, AND OTHER SUGARS IN GLYCOCONJUGATES USING AVIDIN BIOTIN TECHNOLOGY [J].

BAYER, EA ;

BENHUR, H ;

WILCHEK, M .

ANALYTICAL BIOCHEMISTRY, 1988, 170 (02) :271-281

[2] Functional RNA microarrays for high-throughput screening of antiprotein aptamers [J].

Collett, JR ;

Cho, EJ ;

Lee, JF ;

Levy, M ;

Hood, AJ ;

Wan, C ;

Ellington, AD .

ANALYTICAL BIOCHEMISTRY, 2005, 338 (01) :113-123

[3] Unnatural base pairs mediate the site-specific incorporation of an unnatural hydrophobic component into RNA transcripts [J].

Endo, M ;

Mitsui, T ;

Okuni, T ;

Kimoto, A ;

Hirao, I ;

Yokoyama, S .

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2004, 14 (10) :2593-2596

[4] Synthesis of 6-(2-thienyl)purine nucleoside derivatives that form unnatural base pairs with pyridin-2-one nucleosides [J].

Fujiwara, T ;

Kimoto, M ;

Sugiyama, H ;

Hirao, I ;

Yokoyama, S .

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2001, 11 (16) :2221-2223

[5] An unnatural base pair for incorporating amino acid analogs into proteins [J].

Hirao, I ;

Ohtsuki, T ;

Fujiwara, T ;

Mitsui, T ;

Yokogawa, T ;

Okuni, T ;

Nakayama, H ;

Takio, K ;

Yabuki, T ;

Kigawa, T ;

Kodama, K ;

Yokogawa, T ;

Nishikawa, K ;

Yokoyama, S .

NATURE BIOTECHNOLOGY, 2002, 20 (02) :177-182

[6] Synthesis of adenosine derivatives as transcription initiators and preparation of 5′ fluorescein- and biotin-labeled RNA through one-step in vitro transcription [J].

Huang, FQ ;

Wang, GC ;

Coleman, T ;

Li, N .

RNA, 2003, 9 (12) :1562-1570

[7] Site-specific incorporation of a photo-crosslinking component into RNA by T7 transcription mediated by unnatural base pairs [J].

Kimoto, M ;

Endo, M ;

Mitsui, T ;

Okuni, T ;

Hirao, I ;

Yokoyama, S .

CHEMISTRY & BIOLOGY, 2004, 11 (01) :47-55

[8] Anti-(Raf-1) RNA aptamers that inhibit Ras-induced Raf-1 activation [J].

Kimoto, M ;

Shirouzu, M ;

Mizutani, S ;

Koide, H ;

Kaziro, Y ;

Hirao, I ;

Yokoyama, S .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2002, 269 (02) :697-704

[9] ENZYMATIC-SYNTHESIS OF BIOTIN-LABELED POLYNUCLEOTIDES - NOVEL NUCLEIC-ACID AFFINITY PROBES [J].

LANGER, PR ;

WALDROP, AA ;

WARD, DC .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1981, 78 (11) :6633-6637

[10] RAPID AND EFFICIENT SYNTHESIS OF NUCLEOSIDE 5'-O-(1-THIOTRIPHOSPHATES), 5'-TRIPHOSPHATES AND 2',3'-CYCLOPHOSPHOROTHIOATES USING 2-CHLORO-4H-1,3,2-BENZODIOXAPHOSPHORIN-4-ONE [J].

LUDWIG, J ;

ECKSTEIN, F .

JOURNAL OF ORGANIC CHEMISTRY, 1989, 54 (03) :631-635