ArticleNucleic acids research2021
Characterizing and circumventing sequence restrictions for synthesis of circular RNA in vitro.
Article in Nucleic acids research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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The trial behind it
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Who cites it
40 citing papers in PubMed.
- Emerging strategies and innovations in circular RNA synthesis.Molecular therapy. Nucleic acids · 2026Review
- A scalable platform for dumbbell siRNA production using engineeredMolecular therapy. Nucleic acids · 2026Article
- Research Progress, Application, and Industrialization Prospects of Circular RNA Vaccines in Viral Diseases.Vaccines · 2026Review
- Target site selection and P1 engineering enable highly efficient circular RNA production via end-to-end self-targeting and splicing.Nucleic acids research · 2026Article
- Split intron-exon system for circular RNA synthesis.Nature communications · 2026Article
- Review
- Circular RNA-mediated regulation of key signaling pathways in cardiovascular diseases: a review.Journal of advanced research · 2026Review
- Pre-splicing conformation and stepwise circularization of a group I intron in Azoarcus pre-tRNA.Nature communications · 2026Article
- Article
- Impact of PEGylation and hyaluronan functionalization on lipoplex-mediated mRNA delivery to the canine retina.Drug delivery · 2025Article
- Circular RNA: From non-coding regulators to functional protein encoders.Pharmaceutical science advances · 2025Review
- Local delivery of IL-15 and anti-PD-L1 nanobody by in vitro-transcribed circILNb elicits superior antitumor immunity in cold tumors.Cell reports. Medicine · 2025Article
- Development and comprehensive evaluation of scarless circularization systems for circular RNA therapeutics.Molecular therapy. Nucleic acids · 2025Article
- Mechanistic insights into circularization viaMolecular therapy. Nucleic acids · 2025Article
- Exploring the Potential and Advancements of Circular RNA Therapeutics.Exploration (Beijing, China) · 2025Review
- Rational Design and Immunological Mechanisms of Circular RNA-Based Vaccines: Emerging Frontiers in Combating Pathogen Infection.Vaccines · 2025Review
- The therapeutic potential of circular RNAs.Nature reviews. Genetics · 2025Review
- Recent advances and perspectives on the development of circular RNA cancer vaccines.NPJ vaccines · 2025Review
- CircRNA Synthesis and Applications.Advances in experimental medicine and biology · 2025Review
- Review
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Just as eukaryotic circular RNA (circRNA) is a product of intracellular backsplicing, custom circRNA can be synthesized in vitro using a transcription template in which transposed halves of a split group I intron flank the sequence of the RNA to be circularized. Such permuted intron-exon (PIE) constructs have been used to produce circRNA versions of ribozymes, mimics of viral RNA motifs, a streptavidin aptamer, and protein expression vectors for genetic engineering and vaccine development. One limitation of this approach is the obligatory incorporation of small RNA segments (E1 and E2) into nascent circRNA at the site of end-joining. This restriction may preclude synthesis of small circRNA therapeutics and RNA nanoparticles that are sensitive to extraneous sequence, as well as larger circRNA mimics whose sequences must precisely match those of the native species on which they are modelled. In this work, we used serial mutagenesis and in vitro selection to determine how varying E1 and E2 sequences in a thymidylate synthase (td) group I intron PIE transcription template construct affects circRNA synthesis yield. Based on our collective findings, we present guidelines for the design of custom-tailored PIE transcription templates from which synthetic circRNAs of almost any sequence may be efficiently synthesized.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.