Synthesis of N~4-Methylcytidine (m~4C) and N~4,N~4-Dimethylcytidine (m~4_2C) Modified RNA

Ya Ying Zheng; Song Mao; Jia Sheng

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Synthesis of N~4-Methylcytidine (m~4C) and N~4,N~4-Dimethylcytidine (m~4_2C) Modified RNA

机译：Synthesis of N~4-Methylcytidine (m~4C) and N~4,N~4-Dimethylcytidine (m~4_2C) Modified RNA

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This article summarizes the protocols for phosphoramidite chemistry and solid phase synthesis of RNA oligonucleotides containing N~4-methylcytidine (m~4C) and N~4,N~4-dimethylcytidine (m~4_2C) residues for base-pairing, structural, and enzymatic activity studies. The two key m~4C and m~4_2C phosphoramidite building blocks can be synthesized starting from the partially protected cytidine nucleosides, followed by solid-phase synthesis and HPLC purification of the modified target RNA oligonucleotides. These modified RNA strands are then prepared for base pairing stability, specificity, and structural studies using UV-melting temperature (T_m) measurements and X-ray crystallography. Functional studies are performed by reverse transcription assays in primer extension reactions employing different enzymes.

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《Current Protocols in Nucleic Acid Chemistry》 |2021年第9期|e248-1-e248-16|共16页
作者
Ya Ying Zheng; Song Mao; Jia Sheng;
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作者单位

Department of Chemistry and The RNA Institute, University at Albany, State University of New York, Albany, New York;

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正文语种英语
中图分类生物化学;
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base pairing; N~4; N~4-dimethylcytidine (m~4_2C); N~4-methylcytidine (m~4C); nucleic acids modification; thermostability; X-ray crystallography;

Synthesis of N~4-Methylcytidine (m~4C) and N~4,N~4-Dimethylcytidine (m~4_2C) Modified RNA

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