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. 2024 Sep 10;31:89. doi: 10.1186/s12929-024-01080-z

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Synthesis and purification of three distinct mRNA types for mRNA-LNP drugs. A The in vitro transcription (IVT) process is illustrated. First, a plasmid is generated containing the target gene with a T7 promoter. After restriction enzyme (RE) digestion of the plasmid and purification of linear DNA, IVT is performed with T7 RNA polymerase, cap analogue, modified bases, and RNase inhibitor to generate transcribed linear mRNAs. The linear mRNAs may be traditional linear mRNAs, self-amplified mRNAs (saRNAs), or trans-amplified mRNAs (taRNAs). For production of circular RNAs (circRNAs), cyclization is achieved via intron-splicing reaction or T4 RNA ligase. Impurities within the mRNA products may be eliminated by DNA digestion and mRNA purification, along with other methods specific to the type of RNA product. The highly purified mRNAs are suitable for incorporation into mRNA-LNP formulations. (SEC: size exclusion chromatography, HIC: hydrophobic interaction chromatography, RP-HPLC: reverse phase HPLC). B Advantages and disadvantages of the three different mRNA types used for mRNA-LNP drugs are shown