Communication
ChemComm
13 Z. Li, T. Lavergne, D. A. Malyshev, J. Zimmermann, R. Adhikary,
K. Dhami, P. Ordoukhanian, Z. Sun, J. Xiang and F. E. Romesberg,
Chemistry, 2013, 19, 14205–14209; D. A. Malyshev, D. A. Pfaff, S. I.
Ippoliti, G. T. Hwang, T. J. Dwyer and F. E. Romesberg, Chemistry,
2010, 16, 12650–12659.
14 M. Endo, T. Mitsui, T. Okuni, M. Kimoto, I. Hirao and S. Yokoyama,
Bioorg. Med. Chem. Lett., 2004, 14, 2593–2596; M. Kimoto, R. Kawai,
T. Mitsui, S. Yokoyama and I. Hirao, Nucleic Acids Symp. Ser., 2008,
457–458; M. Kimoto, T. Mitsui, S. Yokoyama and I. Hirao, J. Am. Chem.
Soc., 2010, 132, 4988–4989; T. Mitsui, M. Kimoto, Y. Harada, A. Sato,
A. Kitamura, T. To, I. Hirao and S. Yokoyama, Nucleic Acids Res. Suppl.,
2002, 219–220.
15 Y. Hikida, M. Kimoto, S. Yokoyama and I. Hirao, Nat. Protoc., 2010,
5, 1312–1323.
16 T. Mitsui, M. Kimoto, Y. Harada, S. Yokoyama and I. Hirao, J. Am.
Chem. Soc., 2005, 127, 8652–8658.
17 T. Ohtsuki, M. Kimoto, M. Ishikawa, T. Mitsui, I. Hirao and
S. Yokoyama, Proc. Natl. Acad. Sci. U. S. A., 2001, 98, 4922–4925;
R. Kawai, M. Kimoto, S. Ikeda, T. Mitsui, M. Endo, S. Yokoyama and
I. Hirao, J. Am. Chem. Soc., 2005, 127, 17286–17295; N. Morohashi,
M. Kimoto, A. Sato, R. Kawai and I. Hirao, Molecules, 2012, 17,
2855–2876.
addition of functional groups such as fluorophores to RNA with
the selectivity of an unnatural base pair for the introduction of
strained alkenes during transcription, this method is a valuable
tool for the post-transcriptional functionalization of enzymatically
prepared RNA under native conditions. As we have shown, only
full length transcripts are obtained, if the unnatural triphosphate
is present, facilitating downstream applications of the function-
alized RNA. The native structure of the RNA will not be affected by
the mild click reaction conditions, which can be performed in
buffered solutions, water or in a cellular context.23,24,27 This is
essential for the study of RNA constructs with complex folding
pathways such as for example various ribozymes and aptamers.
The presented norbornene building block for fast and copper-free
click reactions might additionally prove useful for site-specific
labeling of specific RNA molecules in cells.
This work was supported by the Fonds der Chemischen Industrie
(Liebig-Fellowship to S.K-S.). We thank M. Famulok (LIMES Institute,
University of Bonn, Germany) for laboratory facilities.
18 T. Ishizuka, M. Kimoto, A. Sato and I. Hirao, Chem. Commun., 2012,
48, 10835–10837.
19 M. Kimoto and I. Hirao, Methods Mol. Biol., 2010, 634, 355–369;
M. Kimoto, R. Yamashige, S. Yokoyama and I. Hirao, J. Nucleic Acids,
2012, 2012, 230943.
Notes and references
20 Y. J. Seo, S. Matsuda and F. E. Romesberg, J. Am. Chem. Soc., 2009,
131, 5046–5047.
21 Y. J. Seo, D. A. Malyshev, T. Lavergne, P. Ordoukhanian and F. E.
Romesberg, J. Am. Chem. Soc., 2011, 133, 19878–19888.
22 M. L. Blackman, M. Royzen and J. M. Fox, J. Am. Chem. Soc., 2008,
130, 13518–13519.
1 E. Paredes, M. Evans and S. R. Das, Methods, 2011, 54, 251–259.
2 I. Hirao, M. Kimoto, T. Mitsui, T. Fujiwara, R. Kawai, A. Sato,
Y. Harada and S. Yokoyama, Nat. Methods, 2006, 3, 729–735.
3 R. Yamashige, M. Kimoto, Y. Takezawa, A. Sato, T. Mitsui, S. Yokoyama
and I. Hirao, Nucleic Acids Res., 2012, 40, 2793–2806.
4 I. Hirao, T. Mitsui, M. Kimoto and S. Yokoyama, J. Am. Chem. Soc.,
2007, 129, 15549–15555; I. Hirao, M. Kimoto and R. Yamashige, Acc.
Chem. Res., 2012, 45, 2055–2065; I. Hirao, T. Ohtsuki, T. Fujiwara,
T. Mitsui, T. Yokogawa, T. Okuni, H. Nakayama, K. Takio, T. Yabuki,
T. Kigawa, K. Kodama, T. Yokogawa, K. Nishikawa and S. Yokoyama,
Nat. Biotechnol., 2002, 20, 177–182.
5 D. A. Malyshev, Y. J. Seo, P. Ordoukhanian and F. E. Romesberg, J. Am.
Chem. Soc., 2009, 131, 14620–14621; K. Betz, D. A. Malyshev, T. Lavergne,
W. Welte, K. Diederichs, T. J. Dwyer, P. Ordoukhanian, F. E. Romesberg
and A. Marx, Nat. Chem. Biol., 2012, 8, 612–614; D. A. Malyshev, K. Dhami,
H. T. Quach, T. Lavergne, P. Ordoukhanian, A. Torkamani and F. E.
Romesberg, Proc. Natl. Acad. Sci. U. S. A., 2012, 109, 12005–12010; K. Betz,
D. A. Malyshev, T. Lavergne, W. Welte, K. Diederichs, F. E. Romesberg and
A. Marx, J. Am. Chem. Soc., 2013, 135, 18637–18643; T. Lavergne,
M. Degardin, D. A. Malyshev, H. T. Quach, K. Dhami, P. Ordoukhanian
and F. E. Romesberg, J. Am. Chem. Soc., 2013, 135, 5408–5419.
23 N. K. Devaraj, R. Weissleder and S. A. Hilderbrand, Bioconjugate
Chem., 2008, 19, 2297–2299.
24 N. K. Devaraj, R. Upadhyay, J. B. Haun, S. A. Hilderbrand and
R. Weissleder, Angew. Chem., Int. Ed., 2009, 48, 7013–7016; D. Schulz
and A. Rentmeister, ChemBioChem, 2014, 15, 2342–2347.
25 E. M. Sletten and C. R. Bertozzi, Angew. Chem., Int. Ed., 2009, 48,
6974–6998; N. K. Devaraj, S. Hilderbrand, R. Upadhyay, R. Mazitschek
and R. Weissleder, Angew. Chem., Int. Ed., 2010, 49, 2869–2872; Z. Li,
H. Cai, M. Hassink, M. L. Blackman, R. C. Brown, P. S. Conti and
J. M. Fox, Chem. Commun., 2010, 46, 8043–8045; N. K. Devaraj and
R. Weissleder, Acc. Chem. Res., 2011, 44, 816–827; K. Lang, L. Davis,
J. Torres-Kolbus, C. Chou, A. Deiters and J. W. Chin, Nat. Chem., 2012, 4,
298–304; D. S. Liu, A. Tangpeerachaikul, R. Selvaraj, M. T. Taylor, J. M.
Fox and A. Y. Ting, J. Am. Chem. Soc., 2012, 134, 792–795; J. L. Seitchik,
J. C. Peeler, M. T. Taylor, M. L. Blackman, T. W. Rhoads, R. B. Cooley,
C. Refakis, J. M. Fox and R. A. Mehl, J. Am. Chem. Soc., 2012, 134,
2898–2901; S. Schneider, M. J. Gattner, M. Vrabel, V. Flugel, V. Lopez-
Carrillo, S. Prill and T. Carell, ChemBioChem, 2013; K. Wang, D. Wang,
K. Ji, W. Chen, Y. Zheng, C. Dai and B. Wang, Org. Biomol. Chem., 2015,
13, 909–915; U. Rieder and N. W. Luedtke, Angew. Chem., Int. Ed., 2014,
53, 9168–9172; J. M. Holstein, D. Stummer and A. Rentmeister, Chem.
Sci., 2015, 6, 1362–1369.
6 Y. J. Seo, G. T. Hwang, P. Ordoukhanian and F. E. Romesberg, J. Am.
Chem. Soc., 2009, 131, 3246–3252.
7 K. Dhami, D. A. Malyshev, P. Ordoukhanian, T. Kubelka, M. Hocek
and F. E. Romesberg, Nucleic Acids Res., 2014, 42, 10235–10244.
8 D. A. Malyshev, K. Dhami, T. Lavergne, T. Chen, N. Dai, J. M. Foster,
I. R. Correa, Jr. and F. E. Romesberg, Nature, 2014, 509, 385–388.
9 C. Switzer, S. E. Moroney and S. A. Benner, J. Am. Chem. Soc., 1989, 111,
8322–8323; C. Y. Switzer, S. E. Moroney and S. A. Benner, Biochemistry, 26 M. R. Karver, R. Weissleder and S. A. Hilderbrand, Bioconjugate Chem.,
1993, 32, 10489–10496; C. R. Geyer, T. R. Battersby and S. A. Benner,
2011, 22, 2263–2270.
Structure, 2003, 11, 1485–1498; M. J. Moser, D. J. Marshall, J. K. Grenier, 27 A. M. Pyka, C. Domnick, F. Braun and S. Kath-Schorr, Bioconjugate
C. D. Kieffer, A. A. Killeen, J. L. Ptacin, C. S. Richmond, E. B. Roesch,
Chem., 2014, 25, 1438–1443.
C. W. Scherrer, C. B. Sherrill, C. V. Van Hout, S. J. Zanton and J. R. 28 D. M. Chadalavada, E. A. Gratton and P. C. Bevilacqua, Biochemistry,
Prudent, Clin. Chem., 2003, 49, 407–414; S. C. Johnson, C. B. Sherrill,
2010, 49, 5321–5330.
D. J. Marshall, M. J. Moser and J. R. Prudent, Nucleic Acids Res., 2004, 32, 29 J. S. New, W. L. Christopher, J. P. Yevich, R. Butler, R. F. Schlemmer, Jr.,
1937–1941; Z. Yang, A. M. Sismour, P. Sheng, N. L. Puskar and
S. A. Benner, Nucleic Acids Res., 2007, 35, 4238–4249; Z. Yang, F. Chen,
C. P. VanderMaelen and J. A. Cipollina, J. Med. Chem., 1989, 32,
1147–1156.
¨ggen, J. Org. Chem., 1976, 41, 2084–2086;
H. Vorbru
¨ggen and B. Bennua, Chem. Ber., 1981, 114, 1279–1286;
H. Vorbrueggen, Acc. Chem. Res., 1995, 28, 509–520.
31 K. Gutsmiedl, C. T. Wirges, V. Ehmke and T. Carell, Org. Lett., 2009,
11, 2405–2408.
J. B. Alvarado and S. A. Benner, J. Am. Chem. Soc., 2011, 133, 15105–15112. 30 U. Niedballa and H. Vorbru
10 M. Kimoto, R. Kawai, T. Mitsui, S. Yokoyama and I. Hirao, Nucleic
Acids Res., 2009, 37, e14; I. Hirao and M. Kimoto, Proc. Jpn. Acad.,
Ser. B, 2012, 88, 345–367.
11 L. Li, M. Degardin, T. Lavergne, D. A. Malyshev, K. Dhami,
P. Ordoukhanian and F. E. Romesberg, J. Am. Chem. Soc., 2014, 32 S. G. Srivatsan and Y. Tor, Nat. Protoc., 2007, 2, 1547–1555.
¨
136, 826–829.
33 J. Schoch, M. Wiessler and A. Jaschke, J. Am. Chem. Soc., 2010, 132,
12 T. J. Matray and E. T. Kool, J. Am. Chem. Soc., 1998, 120, 6191–6192;
T. J. Matray and E. T. Kool, Nature, 1999, 399, 704–708; J. C. Morales
and E. T. Kool, Biochemistry, 2000, 39, 2626–2632.
8846–8847; M. Vrabel, P. Kolle, K. M. Brunner, M. J. Gattner,
V. Lopez-Carrillo, R. de Vivie-Riedle and T. Carell, Chemistry, 2013,
19, 13309–13312.
8256 | Chem. Commun., 2015, 51, 8253--8256
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