The 5’-Hexynyl modifier can be used to incorporate an active alkyne onto the 5’-end of an oligonucleotide. The alkyne is separated from the 5’-end nucleotide base by a 4-carbon spacer arm, which serves to reduce steric interaction between the reactive group and the oligo. The presence of the alkyne allows the user to use “Click Chemistry” (a [3+2] cycloaddition reaction between alkynes and azides, using copper (I) iodide as a catalyst) to conjugate it to a variety of azide-containing labels/tags (e.g., fluorescent dyes, biotin), or oligos, with extremely high regioselectivity and efficiency (1,2). When conjugation to an azide-oligo is desired, preparation of the azide-oligo can be achieved using either an Azidobutyrate NHS Ester or the 5’-Bromohexyl modifier (see their respective tech sheets for details). Click chemistry can be used to form short, cyclic oligos that can be used as research tools in various biophysical and biological studies (3). In particular, they have considerable potential for in vivo work, as cyclic oligos are known to be very stable in serum for up to several days.
1. Huisgen, R. Angew. Chem. Int. Ed. (1963), 2: 565-568.
2. Rostovtsev, V.V., Green, L.G., Fokin, V.V., Sharpless, K.B. A Stepwise Huisgen Cycloaddition Process: Copper(I)-Catalyzed Regioselective Ligation of Azides and Terminal Alkynes. Angew. Chem. Int. Ed. (2002), 41: 2596-2599.
3. Kumar, R., El-Sagheer, A., Tumpane, J., Lincoln, P., Wilhelmsson, L.M., Brown, T. Template-Directed Oligonucleotide Strand Ligation, Covalent Intramolecular DNA Circularization and Catenation Using Click Chemistry. J. Am. Chem. Soc. (2007), 129: 6859-6864.