ArticleNucleic acids research2026
Synthesis of long and functionally active RNAs facilitated by acetal levulinic ester chemistry.
Article in Nucleic acids research, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- Improved Purification of sgRNA and pegRNA by Use of a 5'-Terminal Dioctyloxytrityl (DOT) Protecting Group.The Journal of organic chemistry · 2026Article
- 2'-O-Acetal Levulinic Ester Ribonucleoside 3'-Phosphoramidites for the Solid-Phase Synthesis of Long RNA.Current protocols · 2026Article
- IVT-free, chemically synthesized protein-encoding RNA oligonucleotides for rapid production of personalized cancer vaccines.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Solid Phase Chemical Synthesis of RNA.Methods in molecular biology (Clifton, N.J.) · 2026Article
Corrections and comments
- Erratum issued
Authors and funding
9 authors.
Funding
Abstract
Recent advances in RNA-based therapeutics have created a demand for synthetic RNAs that are 100 nucleotides (nts) or longer. In this study, we present the use of 2'-acetal levulinic ester (2'-ALE) phosphoramidites for the synthesis of long RNAs that are at least 215 nts in length. We have developed protocols for rapid (2-4 min) and efficient coupling (>99%) of 2'-ALE monomers and established a rapid, on-column deprotection of RNA strands requiring short alkylamine treatments at room temperature. The results of these studies enabled the successful syntheses of sgRNAs (99 nt), sgRNA tagged with fluorogenic Mango II and Broccoli aptamers (130-170 nt), and 5'-capped minimal mRNAs (200-215 nt), each exhibiting robust functional activity in both cell-free and cellular systems. We also found that the incorporation of 2'-O-methyl-adenosine in the poly(A) tail of synthetic mRNAs markedly enhanced protein expression, highlighting the ALE platform's compatibility for systematic exploration of RNA chemical diversity. Collectively, these results establish 2'-ALE chemistry as a promising platform for the synthesis of long and functionally active RNAs.
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Registered trials
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