Site-Specific One-Pot Dual Labeling of DNA by Orthogonal Cycloaddition Chemistry

被引：98

作者：

Schoch, Juliane ^{[1
]}

Staudt, Markus ^{[1
]}

Samanta, Ayan ^{[1
]}

Wiessler, Manfred ^{[2
]}

Jaeschke, Andres ^{[1
]}

机构：

[1] Heidelberg Univ, Inst Pharm & Mol Biotechnol, D-69120 Heidelberg, Germany

[2] German Canc Res Ctr, Div Med Phys Oncol E020, D-69120 Heidelberg, Germany

来源：

BIOCONJUGATE CHEMISTRY | 2012年 / 23卷 / 07期

关键词：

CLICK CHEMISTRY; SOLID-PHASE; FUNCTIONALIZATION; BIOCONJUGATION; RNA; DIENOPHILES; LIGATION;

D O I：

10.1021/bc300181n

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Bioorthogonal reactions are of high interest in biosciences as they allow the introduction of fluorescent dyes, affinity tags, or other unnatural moieties into biomolecules. The site-specific attachment of two or more different labels is particularly demanding and typically requires laborious multistep syntheses. Here, we report that the most popular cycloaddition in bioconjugation, the copper-catalyzed azide-alkyne click reaction (CuAAC), is fully orthogonal to the inverse electron-demand Diels-Alder reaction (DAinv). We demonstrate that both bioorthogonal reactions can be conducted concurrently in a one pot reaction by just mixing all components. Orthogonality has been established even for highly reactive trans-cyclooctene-based dienophiles (with rate constants up to 380 000 M-1 s(-1)). These properties allow for the convenient site specific one-step preparation of oligonucleotide FRET probes and related reporters needed in cellular biology and biophysical chemistry.

引用

页码：1382 / 1386

页数：5

共 38 条

[1] Chemical Synthesis of Site-Specifically 2′-Azido-Modified RNA and Potential Applications for Bioconjugation and RNA Interference [J].

Aigner, Michaela ;

Hartl, Markus ;

Fauster, Katja ;

Steger, Jessica ;

Bister, Klaus ;

Micura, Ronald .

CHEMBIOCHEM, 2011, 12 (01) :47-51

[2]

Beaucage Serge L., 1993, V20, P33

[3] Click Chemistry and Bioorthogonal Reactions: Unprecedented Selectivity in the Labeling of Biological Molecules [J].

Best, Michael D. .

BIOCHEMISTRY, 2009, 48 (28) :6571-6584

[4] Tetrazine ligation: Fast bioconjugation based on inverse-electron-demand Diels-Alder reactivity [J].

Blackman, Melissa L. ;

Royzen, Maksim ;

Fox, Joseph M. .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2008, 130 (41) :13518-+

[5] Clicking 1,2,4,5-tetrazine and cyclooctynes with tunable reaction rates [J].

Chen, Weixuan ;

Wang, Danzhu ;

Dai, Chaofeng ;

Hamelberg, Donald ;

Wang, Binghe .

CHEMICAL COMMUNICATIONS, 2012, 48 (12) :1736-1738

[6] Bioconjugation with Strained Alkenes and Alkynes [J].

Debets, Marjoke F. ;

Van Berkel, Sander S. ;

Dommerholt, Jan ;

Dirks, A. J. ;

Rutjes, Floris P. J. T. ;

Van Delft, Floris L. .

ACCOUNTS OF CHEMICAL RESEARCH, 2011, 44 (09) :805-815

[7] Biocompatible Copper(I) Catalysts for in Vivo Imaging of Glycans [J].

del Amo, David Soriano ;

Wang, Wei ;

Jiang, Hao ;

Besanceney, Christen ;

Yan, Amy C. ;

Levy, Matthew ;

Liu, Yi ;

Marlow, Florence L. ;

Wu, Peng .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2010, 132 (47) :16893-16899

[8] Biomedical Applications of Tetrazine Cycloadditions [J].

Devaraj, Neal K. ;

Weissleder, Ralph .

ACCOUNTS OF CHEMICAL RESEARCH, 2011, 44 (09) :816-827

[9] Click chemistry with DNA [J].

El-Sagheer, Afaf H. ;

Brown, Tom .

CHEMICAL SOCIETY REVIEWS, 2010, 39 (04) :1388-1405

[10] 2′-Azido RNA, a Versatile Tool for Chemical Biology: Synthesis, X-ray Structure, siRNA Applications, Click Labeling [J].

Fauster, Katja ;

Hartl, Markus ;

Santner, Tobias ;

Aigner, Michaela ;

Kreutz, Christoph ;

Bister, Klaus ;

Ennifar, Eric ;

Micura, Ronald .

ACS CHEMICAL BIOLOGY, 2012, 7 (03) :581-589

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