Modified Nucleic Acid for Systematic Evolution of RNA Ligands by Exponential Enrichment

Abstract

Various types of modified nucleic acid were tested for systematic evolution of ligands by exponential enrichment (SELEX) or in vitro evolution. T7 RNA polymerase accepted cytidine triphosphate with a biotinyl group at the N⁴-position of cytosine, adenosine triphosphate with an aminohexyl group at the N⁶-position of adenine, and 2T-amino-2′-deoxyuridine triphosphate as substrates for transcription of DNA containing a random sequence. However, uridine triphosphate with a nitroveratryloxy group at the 2′-position and adenosine triphosphate with an aminohexyl group at the 8-position of adenine were not accepted. The transcribed products served as templates for the production of cDNA by reverse transcription. These findings indicate that some modified nucleotides can be applied to the present SELEX protocol.

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References

1. Ellington, A. D. and Szostak, J. W. 1990. Nature, 346:818.

2. Tuerk, C. and Gold, L. 1990. Science, 249:505.

3. Kawazoe, N. , Ito, Y. and Imanishi, Y. 1996. Anal. Chem., 68:4309.

4. Ito, Y. , Suzuki, A. , Kawazoe, N. and Imanishi, Y. 1997. Bull. Chem. Soc. Jpn., 70:695.

5. Pomerantz, S. C. , McCloskey, J. A. , Tarasow, T. M. and Eaton, B. E. 1997. J. Am. Chem. Soc., 119:3861.

6. Eaton, B. E. and Pieken, W. A. 1995. Annu. Rev. Biochem., 64:837.

7. Eaton, B. E. 1997. Curr. Opin. Chem. Biol., 1:10.

8. Lin, Y. , Qiu, Q. , Gill, S. C. and Jayasena, S. D. 1994. Nucleic Acids Res., 22:5229.

9. Lin, Y. , Nieuwlandt, D. , Magallanez, A. , Feistner, B. and Jayasena, S. D. 1996. Nucleic. Acids Res., 24:3407.

10.

10. Lee, S. and Sullenger, B. A. 1997. Nature Biotechnology, 15:41.

11.

11. Pagratis, N. C. , Bell, C. , Chang, Y. , Jennings, S. , Fitzwater, T. , Jellinek, D. and Dang, C. 1997. Nature Biotechnology, 15:68.

12.

12. Pieken, W. A. , Olsen, D. B. , Benseler, F. , Aurup, H. and Eckstein, F. 1991. Science, 253:314.

13.

13. Jensen, K. B. , Atkinson, B. L. , Willis, M. C. , Koch, T. H. and Gold, L. 1995. Proc. Natl. Acad. Sci. US.A., 92:12220.

14.

14. Dewey, T. M. , Mundt, A. A. , Crouch, G. J. , Zyzniewski, M. C. and Eaton, B. E. 1995. J. Am. Chem. Soc., 117:8474.

15.

15. Tarasow, T. M. , Tarasow, S. L. and Eaton, B. E. 1997. Nature, 389:54.

16.

16. Conrad, E. , Hanne, A. , Gaur, R. K. and Krupp, G. 1995. Nucleic Acids Res., 23:1845.