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Synthesis and characterization of 2'-modified-4'-thioRNA: a comprehensive comparison of nuclease stability.

Takahashi M, Minakawa N, Matsuda A - Nucleic Acids Res. (2009)

Bottom Line: The results of a comprehensive comparison of nuclease stability of single-stranded F-SRNA and Me-SRNA along with 2'-fluoroRNA (FRNA), 2'-O-MeRNA (MeRNA), SRNA, and natural RNA and DNA, revealed that Me-SRNA had the highest stability with t(1/2) values of > 24 h against S1 nuclease (an endonuclease) and 79.2 min against SVPD (a 3'-exonuclease).Moreover, the stability of Me-SRNA was significantly improved in 50% human plasma (t(1/2) = 1631 min) compared with FRNA (t(1/2) = 53.2 min) and MeRNA (t(1/2) = 187 min), whose modifications are currently used as components of therapeutic aptamers.The results presented in this article will, it is hoped, contribute to the development of 2'-modified-4'-thioRNAs, especially Me-SRNA, as a new RNA molecule for therapeutic applications.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.

ABSTRACT
We report herein the synthesis and physical and physiological characterization of fully modified 2'-modified-4'-thioRNAs, i.e. 2'-fluoro-4'-thioRNA (F-SRNA) and 2'-O-Me-4'-thioRNA (Me-SRNA), which can be considered as a hybrid chemical modification based on 2'-modified oligonucleotides (ONs) and 4'-thioRNA (SRNA). In its hybridization with a complementary RNA, F-SRNA (15mer) showed the highest T(m) value (+16 degrees C relative to the natural RNA duplex). In addition, both F-SRNA and Me-SRNA preferred RNA as a complementary partner rather than DNA in duplex formation. The results of a comprehensive comparison of nuclease stability of single-stranded F-SRNA and Me-SRNA along with 2'-fluoroRNA (FRNA), 2'-O-MeRNA (MeRNA), SRNA, and natural RNA and DNA, revealed that Me-SRNA had the highest stability with t(1/2) values of > 24 h against S1 nuclease (an endonuclease) and 79.2 min against SVPD (a 3'-exonuclease). Moreover, the stability of Me-SRNA was significantly improved in 50% human plasma (t(1/2) = 1631 min) compared with FRNA (t(1/2) = 53.2 min) and MeRNA (t(1/2) = 187 min), whose modifications are currently used as components of therapeutic aptamers. The results presented in this article will, it is hoped, contribute to the development of 2'-modified-4'-thioRNAs, especially Me-SRNA, as a new RNA molecule for therapeutic applications.

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Mentions: Synthetic protocols shown in Schemes 1 and 2 were highlighted in this article. Protocols for synthesis of the other units along with CPG support for F-SRNA and Me-SRNA synthesis were given in the Supplementary Data (Schemes S1, S2 and S3).Scheme 1.Scheme 2.


Synthesis and characterization of 2'-modified-4'-thioRNA: a comprehensive comparison of nuclease stability.

Takahashi M, Minakawa N, Matsuda A - Nucleic Acids Res. (2009)

© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651785&req=5

Mentions: Synthetic protocols shown in Schemes 1 and 2 were highlighted in this article. Protocols for synthesis of the other units along with CPG support for F-SRNA and Me-SRNA synthesis were given in the Supplementary Data (Schemes S1, S2 and S3).Scheme 1.Scheme 2.

Bottom Line: The results of a comprehensive comparison of nuclease stability of single-stranded F-SRNA and Me-SRNA along with 2'-fluoroRNA (FRNA), 2'-O-MeRNA (MeRNA), SRNA, and natural RNA and DNA, revealed that Me-SRNA had the highest stability with t(1/2) values of > 24 h against S1 nuclease (an endonuclease) and 79.2 min against SVPD (a 3'-exonuclease).Moreover, the stability of Me-SRNA was significantly improved in 50% human plasma (t(1/2) = 1631 min) compared with FRNA (t(1/2) = 53.2 min) and MeRNA (t(1/2) = 187 min), whose modifications are currently used as components of therapeutic aptamers.The results presented in this article will, it is hoped, contribute to the development of 2'-modified-4'-thioRNAs, especially Me-SRNA, as a new RNA molecule for therapeutic applications.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.

ABSTRACT
We report herein the synthesis and physical and physiological characterization of fully modified 2'-modified-4'-thioRNAs, i.e. 2'-fluoro-4'-thioRNA (F-SRNA) and 2'-O-Me-4'-thioRNA (Me-SRNA), which can be considered as a hybrid chemical modification based on 2'-modified oligonucleotides (ONs) and 4'-thioRNA (SRNA). In its hybridization with a complementary RNA, F-SRNA (15mer) showed the highest T(m) value (+16 degrees C relative to the natural RNA duplex). In addition, both F-SRNA and Me-SRNA preferred RNA as a complementary partner rather than DNA in duplex formation. The results of a comprehensive comparison of nuclease stability of single-stranded F-SRNA and Me-SRNA along with 2'-fluoroRNA (FRNA), 2'-O-MeRNA (MeRNA), SRNA, and natural RNA and DNA, revealed that Me-SRNA had the highest stability with t(1/2) values of > 24 h against S1 nuclease (an endonuclease) and 79.2 min against SVPD (a 3'-exonuclease). Moreover, the stability of Me-SRNA was significantly improved in 50% human plasma (t(1/2) = 1631 min) compared with FRNA (t(1/2) = 53.2 min) and MeRNA (t(1/2) = 187 min), whose modifications are currently used as components of therapeutic aptamers. The results presented in this article will, it is hoped, contribute to the development of 2'-modified-4'-thioRNAs, especially Me-SRNA, as a new RNA molecule for therapeutic applications.

Show MeSH