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Custom RNA Oligonucleotide Synthesis

Gene Link specializes in the synthesis of RNA modified oligos, RNA fluorescent probes with quenchers, chimeric oligos containing various combinations of DNA, 2'O methyl bases and phosphorothioate linkages.

RNA and 2’-O-methyl RNA oligonucleotide synthesis is performed at Gene Link using the b cyanoethyl chemistry and state of the art synthesizers. These involve proprietary software protocols with long coupling times and specialized cycles to obtain ultra clean oligos.

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. The coupling efficiency of 2’-O- methyl phosphoramidite is also higher than the RNA monomers resulting in higher yield of full length oligos.

Gene Link 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 phoshodiester 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 (please refer to our technical sheet on Antisense Oligonucleotides for other modifications). Custom phosphorothioate oligonucleotides synthesized by Gene Link 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. Goodwin,J.T., Stanick,W.A. and Glick,G.D. (1994) Improved solid phase synthesis of long oligoribonucleotides. Application to tRNA (phe) and tRNA(gly). J. Org. Chem. 59:7941-7943
2. Cotton,M., Oberhauser,B., Burnar, H. et al. (1991) 2O methyl and 2O ethyl oligoribonucleotides as inhibitors of the in vitro U7 snRNP-dependent messenger-RNA processing event. NAR 19:2629
3. Milligan, J.F., Matteucci, M.D. and Martin, J.C. (1993) Current concepts in antisense drug design. J. Medicinal Chem. 36:1923-1937.
4. Helene, C., Toulme, J. (1990) Specific regulation of gene expression by antisense, sense and antigene nucleic acids. Biochim. Biophys. Acta. 1049: 99-125.
5. Weintraub, H. M. (1990) Antisense RNA and DNA. Sci. Amer. 262:40-46.
6. Iyer, R.P., Egan. W., Regan, J.B and Beaucage, S.L. (1990) J. Am. Chem. Soc.112; 1253-1254.
7. 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.

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