RNA Oligos RNA Degradation 2’-O- Methyl Phosphoramidite

RNA oligos are susceptible to degradation to the same extent as native RNA extracted from various sources. An attractive alternate to prevent degradation from nucleases is the use of 2’-O- methyl RNA bases, when specific 2’OH is not required. The 2’-O- methyl oligonucleotides confer considerable nuclease resistance and are similar in hydrogen bonding properties to RNA/RNA than the lower RNA/DNA binding property (7). The coupling efficiency of 2’-O- methyl phosphoramidite is also higher than the RNA monomers resulting in higher yield of full-length oligos. We also offers custom synthesis of RNA and DNA chimeric oligos with investigator specified ribo or deoxy bases or 2’-O-methyl bases. The chimeric oligos can also be synthesized with the regular phosphodiester bonds or substituted with phosphorothioate linkages. The combination of 2’-O- methyl RNA bases with phosphorothioate internucleotide linkages imparts these oligos greater nuclease resistance, which is particularly useful for antisense studies. Custom phosphorothioate oligonucleotides synthesized by us can be specified to have all the diester bonds substituted or only some selected diester linkages depending upon the researchers experimental requirement. Substitution of all diester linkage is recommended to provide greater nuclease resistance.

References

1. Milligan, J.F., Matteucci, M.D. and Martin, J.C. (1993) Current concepts in antisense drug design. J. Medicinal Chem. 36:1923-1937.
2. Helene, C., Toulme, J. (1990) Specific regulation of gene expression by antisense, sense and antigen nucleic acids. Biochem. Biophys. Acta. 1049: 99-125.
3. Weintraub, H. M. (1990) Antisense RNA and DNA. Sci. Amer. 262:40-46.
4. Iyer, R.P., Egan. W., Regan, J.B and Beaucage, S.L. (1990) J. Am. Chem. Soc.112; 1253-1254.
5. Wagner, R.W., Matteucci, M.D., Lewis, J.G., Gutierrez, A.J., Moulds, C. and Froehler, B.C. (1993) Antisense gene inhibition by oligonucleotides containing C-5 propyne pyrimidines. Science 260:1510-1513.
6. Hertoghs, K.M.L., Ellis, J.H. and Catchpole, I.R (2003) Use of locked nucleic acid oligonucleotides to add functionality to plasmid DNA. Nucl. Acids Res.31 (20): 5817-5830.
7. Cotton, M., Oberhauser, B., Burnar, H. et al. (1991) 2’O methyl and 2’O ethyl oligoribonucleotides as inhibitors of the in vitro U7 snRNP-dependent messenger-RNA processing event. NAR 19:2629.
8. Singh, S.K., Nielsen, P., Koshkin, A.A. and J. Wengel, Chem. Comm., 1998, (4), 455-456.
9. A.A. Koshkin, A.A., Singh, S.K., Nielsen, P., Rajwanshi, V.K., Kumar, R., Meldgaard, M., Olsen, C.E and Wengel, J. (1998) Tetrahedron 54:3607-3630.
10. Kværnø, L. and Wengel, J. (1999) Chem. Comm., 7:657-658.
11. Petersen, M and Wengel, J. (2003) Trends in Biotechnology 21(2): 74-81.
12. P.A. Giannaris, P.A. and Damha, M.J (1993) Nucleic Acids Research, 21:4742-4749.
13. Bhan, P., Bhan, A., Hong, M.K., Hartwell, J.G., Saunders, J.M and Hoke, G.D (1997) Nucleic Acids Res, 25:3310-3317.
14. Coleman, R.S. and Kesicki, E.S (1994) J. Amer. Chem. Soc., 116:11636-11642. Antisense