Avijit Sahoo, Kiran R. Gore
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{"title":"合成 N3-甲基尿苷和 2'-O 烷基/2'-氟-N3-甲基尿苷修饰的磷酰胺及其与 DNA 和 RNA 寡核苷酸的结合。","authors":"Avijit Sahoo, Kiran R. Gore","doi":"10.1002/cpz1.70039","DOIUrl":null,"url":null,"abstract":"<p>In this article, we describe the synthesis of <i>N</i><sup>3</sup>-methyluridine (m<sup>3</sup>U) and 2′-<i>O</i>-alkyl/2′-fluoro-<i>N</i><sup>3</sup>-methyluridine (2′-<i>O</i>-alkyl/2′-F-m<sup>3</sup>U) phosphoramidites as well as their incorporation into a 14-mer DNA and RNA oligonucleotide sequence. Synthesis of the 2′-<i>O</i>-alkyl-m<sup>3</sup>U phosphoramidite starts with commercially available uridine to achieve a tritylated m<sup>3</sup>U intermediate, followed by 2′-<i>O</i>-alkylation and finally phosphitylation. Synthesis of the 2′-F-m<sup>3</sup>U phosphoramidite is obtained from a commercially available 2′-F-uridine nucleoside. These phosphoramidite monomers are compatible with DNA and RNA oligonucleotide synthesis using conventional phosphoramidite chemistry. This strategy offers efficient synthetic access to various modifications at the 2′-position of m<sup>3</sup>U that can be employed in numerous nucleic acid–based therapeutic applications, including antisense technologies, small interfering RNAs, CRISPR-Cas9, and aptamers. The data presented in this article are based on our previously published reports. © 2024 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Synthesis of 2′-<i>O</i>-alkyl-<i>N</i><sup>3</sup>-methyluridine analogs and their corresponding phosphoramidites</p><p><b>Alternate Protocol 1</b>: Synthesis of 2′-<i>O</i>-TBDMS-<i>N</i><sup>3</sup>-methyluridine and its phosphoramidite</p><p><b>Alternate Protocol 2</b>: Synthesis of 2′-fluoro-<i>N</i><sup>3</sup>-methyluridine and its phosphoramidite</p><p><b>Basic Protocol 2</b>: Solid-phase synthesis of <i>N</i><sup>3</sup>-methyluridine-modified DNA and RNA oligonucleotides</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Synthesis of N3-Methyluridine- and 2′-O-Alkyl/2′-Fluoro-N3-Methyluridine-Modified Phosphoramidites and Their Incorporation into DNA and RNA Oligonucleotides\",\"authors\":\"Avijit Sahoo, Kiran R. Gore\",\"doi\":\"10.1002/cpz1.70039\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>In this article, we describe the synthesis of <i>N</i><sup>3</sup>-methyluridine (m<sup>3</sup>U) and 2′-<i>O</i>-alkyl/2′-fluoro-<i>N</i><sup>3</sup>-methyluridine (2′-<i>O</i>-alkyl/2′-F-m<sup>3</sup>U) phosphoramidites as well as their incorporation into a 14-mer DNA and RNA oligonucleotide sequence. Synthesis of the 2′-<i>O</i>-alkyl-m<sup>3</sup>U phosphoramidite starts with commercially available uridine to achieve a tritylated m<sup>3</sup>U intermediate, followed by 2′-<i>O</i>-alkylation and finally phosphitylation. Synthesis of the 2′-F-m<sup>3</sup>U phosphoramidite is obtained from a commercially available 2′-F-uridine nucleoside. These phosphoramidite monomers are compatible with DNA and RNA oligonucleotide synthesis using conventional phosphoramidite chemistry. This strategy offers efficient synthetic access to various modifications at the 2′-position of m<sup>3</sup>U that can be employed in numerous nucleic acid–based therapeutic applications, including antisense technologies, small interfering RNAs, CRISPR-Cas9, and aptamers. The data presented in this article are based on our previously published reports. © 2024 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Synthesis of 2′-<i>O</i>-alkyl-<i>N</i><sup>3</sup>-methyluridine analogs and their corresponding phosphoramidites</p><p><b>Alternate Protocol 1</b>: Synthesis of 2′-<i>O</i>-TBDMS-<i>N</i><sup>3</sup>-methyluridine and its phosphoramidite</p><p><b>Alternate Protocol 2</b>: Synthesis of 2′-fluoro-<i>N</i><sup>3</sup>-methyluridine and its phosphoramidite</p><p><b>Basic Protocol 2</b>: Solid-phase synthesis of <i>N</i><sup>3</sup>-methyluridine-modified DNA and RNA oligonucleotides</p>\",\"PeriodicalId\":93970,\"journal\":{\"name\":\"Current protocols\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-10-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.70039\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.70039","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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Synthesis of N3-Methyluridine- and 2′-O-Alkyl/2′-Fluoro-N3-Methyluridine-Modified Phosphoramidites and Their Incorporation into DNA and RNA Oligonucleotides
In this article, we describe the synthesis of N 3 -methyluridine (m3 U) and 2′-O -alkyl/2′-fluoro-N 3 -methyluridine (2′-O -alkyl/2′-F-m3 U) phosphoramidites as well as their incorporation into a 14-mer DNA and RNA oligonucleotide sequence. Synthesis of the 2′-O -alkyl-m3 U phosphoramidite starts with commercially available uridine to achieve a tritylated m3 U intermediate, followed by 2′-O -alkylation and finally phosphitylation. Synthesis of the 2′-F-m3 U phosphoramidite is obtained from a commercially available 2′-F-uridine nucleoside. These phosphoramidite monomers are compatible with DNA and RNA oligonucleotide synthesis using conventional phosphoramidite chemistry. This strategy offers efficient synthetic access to various modifications at the 2′-position of m3 U that can be employed in numerous nucleic acid–based therapeutic applications, including antisense technologies, small interfering RNAs, CRISPR-Cas9, and aptamers. The data presented in this article are based on our previously published reports. © 2024 Wiley Periodicals LLC.
Basic Protocol 1 : Synthesis of 2′-O -alkyl-N 3 -methyluridine analogs and their corresponding phosphoramidites
Alternate Protocol 1 : Synthesis of 2′-O -TBDMS-N 3 -methyluridine and its phosphoramidite
Alternate Protocol 2 : Synthesis of 2′-fluoro-N 3 -methyluridine and its phosphoramidite
Basic Protocol 2 : Solid-phase synthesis of N 3 -methyluridine-modified DNA and RNA oligonucleotides