In Vitro Transcribed RNA-Based Platform Vaccines: Past, Present, and Future

mRNA was discovered in 1961, but it was not used as a vaccine until after three decades. Recently, the development of mRNA vaccine technology gained great impetus from the pursuit of vaccines against COVID-19. To improve the properties of RNA vaccines, and primarily their circulation time, self-amplifying mRNA and trans-amplifying mRNA were developed. A separate branch of mRNA technology is circular RNA vaccines, which were developed with the discovery of the possibility of translation on their protein matrix. Circular RNA has several advantages over mRNA vaccines and is considered a fairly promising platform, as is trans-amplifying mRNA. This review presents an overview of the mRNA platform and a critical discussion of the more modern self-amplifying mRNA, trans-amplifying mRNA, and circular RNA platforms created on its basis. Finally, the main features, advantages, and disadvantages of each of the presented mRNA platforms are discussed. This discussion will facilitate the decision-making process in selecting the most appropriate platform for creating RNA vaccines against cancer or viral diseases.

[1]  Seong-Wook Lee,et al.  Efficient circular RNA engineering by end-to-end self-targeting and splicing reaction using Tetrahymena group I intron ribozyme , 2023, Molecular therapy. Nucleic acids.

[2]  M. Nassar,et al.  Recent Advancement in mRNA Vaccine Development and Applications , 2023, Pharmaceutics.

[3]  T. Dzieciątkowski,et al.  mRNA vaccines: The future of prevention of viral infections? , 2023, Journal of medical virology.

[4]  Wahid Khan,et al.  A Comprehensive Review of mRNA Vaccines , 2023, International journal of molecular sciences.

[5]  A. Deviatkin,et al.  Cap-Independent Circular mRNA Translation Efficiency , 2023, Vaccines.

[6]  Qiu Zhao,et al.  Advances of mRNA vaccine in tumor: a maze of opportunities and challenges , 2023, Biomarker Research.

[7]  Zhenglun Liang,et al.  Research progress on circular RNA vaccines , 2023, Frontiers in Immunology.

[8]  U. Şahin,et al.  A trans-amplifying RNA simplified to essential elements is highly replicative and robustly immunogenic in mice. , 2023, Molecular Therapy.

[9]  Y. G. Chen,et al.  Differences in the immunogenicity of engineered circular RNAs , 2023, Journal of molecular cell biology.

[10]  B. Schnierle,et al.  Self-Amplifying RNA Vaccine Candidates: Alternative Platforms for mRNA Vaccine Development , 2023, Pathogens.

[11]  D. Luo,et al.  Targeting the alphavirus virus replication process for antiviral development. , 2022, Antiviral research.

[12]  Shuaicheng Li,et al.  Manganese-coordinated mRNA vaccines with enhanced mRNA expression and immunogenicity induce robust immune responses against SARS-CoV-2 variants , 2022, Science advances.

[13]  E. Settembre,et al.  Self-amplifying mRNA bicistronic influenza vaccines raise cross-reactive immune responses in mice and prevent infection in ferrets , 2022, Molecular therapy. Methods & clinical development.

[14]  U. Şahin,et al.  A Bivalent Trans-Amplifying RNA Vaccine Candidate Induces Potent Chikungunya and Ross River Virus Specific Immune Responses , 2022, Vaccines.

[15]  Howard Y. Chang,et al.  Engineering circular RNA for enhanced protein production , 2022, Nature Biotechnology.

[16]  J. Kulkarni,et al.  The role of lipid components in lipid nanoparticles for vaccines and gene therapy , 2022, Advanced Drug Delivery Reviews.

[17]  A. Rentmeister,et al.  Photocaged 5′ cap analogues for optical control of mRNA translation in cells , 2022, Nature Chemistry.

[18]  Jianxun Song,et al.  mRNA Vaccines: The Dawn of a New Era of Cancer Immunotherapy , 2022, Frontiers in Immunology.

[19]  Yun Yang,et al.  A flexible, efficient, and scalable platform to produce circular RNAs as new therapeutics , 2022, bioRxiv.

[20]  Min Wu,et al.  mRNA-based therapeutics: powerful and versatile tools to combat diseases , 2022, Signal Transduction and Targeted Therapy.

[21]  U. Şahin,et al.  A taRNA vaccine candidate induces a specific immune response that protects mice against Chikungunya virus infections , 2022, Molecular therapy. Nucleic acids.

[22]  J. Neyts,et al.  A dual-antigen self-amplifying RNA SARS-CoV-2 vaccine induces potent humoral and cellular immune responses and protects against SARS-CoV-2 variants through T cell-mediated immunity , 2022, Molecular Therapy.

[23]  Yuhua Li,et al.  Advances in COVID-19 mRNA vaccine development , 2022, Signal Transduction and Targeted Therapy.

[24]  E. Settembre,et al.  Self-amplifying mRNA SARS-CoV-2 vaccines raise cross-reactive immune response to variants and prevent infection in animal models , 2022, Molecular Therapy - Methods & Clinical Development.

[25]  Rui Zhang,et al.  Nonviral Delivery Systems of mRNA Vaccines for Cancer Gene Therapy , 2022, Pharmaceutics.

[26]  D. Cysewski,et al.  2′-O-Methylation of the second transcribed nucleotide within the mRNA 5′ cap impacts the protein production level in a cell-specific manner and contributes to RNA immune evasion , 2022, bioRxiv.

[27]  S. McCormack,et al.  Safety and immunogenicity of a self-amplifying RNA vaccine against COVID-19: COVAC1, a phase I, dose-ranging trial , 2022, eClinicalMedicine.

[28]  Xiaozhong Peng,et al.  Circular RNA vaccines against SARS-CoV-2 and emerging variants , 2022, Cell.

[29]  C. Chakraborty,et al.  Bioengineering of Novel Non-Replicating mRNA (NRM) and Self-Amplifying mRNA (SAM) Vaccine Candidates Against SARS-CoV-2 Using Immunoinformatics Approach , 2022, Molecular Biotechnology.

[30]  M. Gossen,et al.  Chemical modification of uridine modulates mRNA-mediated proinflammatory and antiviral response in primary human macrophages , 2022, Molecular therapy. Nucleic acids.

[31]  S. Biswas,et al.  Demystifying mRNA vaccines: an emerging platform at the forefront of cryptic diseases , 2022, RNA biology.

[32]  N. Shcherbik,et al.  Short and Sweet: Viral 5`-UTR as a Canonical and Non-Canonical Translation Initiation Switch , 2021, Journal of cellular immunology.

[33]  Ling-Ling Chen,et al.  RNA circles with minimized immunogenicity as potent PKR inhibitors. , 2021, Molecular cell.

[34]  Xinjie Chen,et al.  Circular RNA: Biosynthesis in vitro , 2021, Frontiers in Bioengineering and Biotechnology.

[35]  X. Mu,et al.  Immunogenicity of In Vitro-Transcribed RNA. , 2021, Accounts of chemical research.

[36]  B. Verrier,et al.  Combining an optimized mRNA template with a double purification process allows strong expression of in vitro transcribed mRNA , 2021, Molecular Therapy - Nucleic Acids.

[37]  Zhe-Sheng Chen,et al.  RNA methylation and cancer treatment. , 2021, Pharmacological research.

[38]  A. Bazhin,et al.  mRNA-Based Cancer Vaccines: A Therapeutic Strategy for the Treatment of Melanoma Patients , 2021, Vaccines.

[39]  E. Dolgin.  The tangled history of mRNA vaccines , 2021, Nature.

[40]  Yusheng Liu,et al.  Enhancement of synthetic mRNA translation efficiency through engineered poly(A) tails , 2021, bioRxiv.

[41]  Bao-Zhong Wang,et al.  An Update on mRNA-Based Viral Vaccines , 2021, Vaccines.

[42]  R. Langer,et al.  Lipid nanoparticles for mRNA delivery , 2021, Nature Reviews Materials.

[43]  T. Bullock.  CD40 stimulation as a molecular adjuvant for cancer vaccines and other immunotherapies , 2021, Cellular & molecular immunology.

[44]  X. Xia.  Detailed Dissection and Critical Evaluation of the Pfizer/BioNTech and Moderna mRNA Vaccines , 2021, Vaccines.

[45]  Jianzhao Liu,et al.  Synthetic modified messenger RNA for therapeutic applications , 2021, Acta Biomaterialia.

[46]  M. Salehi,et al.  Design of a multi-epitope vaccine against cervical cancer using immunoinformatics approaches , 2021, Scientific Reports.

[47]  W. Cho,et al.  An overview of rational design of mRNA-based therapeutics and vaccines , 2021, Expert opinion on drug discovery.

[48]  J. Meier,et al.  Modifications in an Emergency: The Role of N1-Methylpseudouridine in COVID-19 Vaccines , 2021, ACS central science.

[49]  D. Prazeres,et al.  mRNA vaccines manufacturing: Challenges and bottlenecks , 2021, Vaccine.

[50]  Prisca Obi,et al.  The Design and Synthesis of Circular RNAs. , 2021, Methods.

[51]  S. Pascolo.  Synthetic Messenger RNA-Based Vaccines: From Scorn to Hype , 2021, Viruses.

[52]  M. Houston,et al.  Cap 1 Messenger RNA Synthesis with Co‐transcriptional CleanCap® Analog by In Vitro Transcription , 2021, Current protocols.

[53]  M. Gorospe,et al.  Characterizing and circumventing sequence restrictions for synthesis of circular RNA in vitro. , 2021, Nucleic acids research.

[54]  Jeonghwan Kim,et al.  Self-assembled mRNA vaccines , 2021, Advanced Drug Delivery Reviews.

[55]  D. Glebe,et al.  Host-cell interactions in HBV infection and pathogenesis: the emerging role of m6A modification , 2021, Emerging microbes & infections.

[56]  Trinh T. Tat,et al.  From m6A to Cap-Adjacent m6Am and their Effects on mRNAs , 2020, Epitranscriptomics.

[57]  K. Bloom,et al.  Self-amplifying RNA vaccines for infectious diseases , 2020, Gene Therapy.

[58]  Charles Y. Tan,et al.  A prefusion SARS-CoV-2 spike RNA vaccine is highly immunogenic and prevents lung infection in non-human primates , 2020, bioRxiv.

[59]  Lu Zhang,et al.  mRNA Vaccine Era—Mechanisms, Drug Platform and Clinical Prospection , 2020, International journal of molecular sciences.

[60]  Lufei Wang,et al.  AIM2 Inflammasome's First Decade of Discovery: Focus on Oral Diseases , 2020, Frontiers in Immunology.

[61]  Jessica L. Cohen,et al.  Development of a potent Zika virus vaccine using self-amplifying messenger RNA , 2020, Science Advances.

[62]  J. Yewdell,et al.  Decoding mRNA translatability and stability from the 5′ UTR , 2020, Nature Structural & Molecular Biology.

[63]  K. Majumder,et al.  Immuno-informatics approach for multi-epitope vaccine designing against SARS-CoV-2 , 2020, bioRxiv.

[64]  Shahin Nazarian,et al.  An in silico deep learning approach to multi-epitope vaccine design: a SARS-CoV-2 case study , 2020, Scientific Reports.

[65]  H. Feldmann,et al.  An alphavirus-derived replicon RNA vaccine induces SARS-CoV-2 neutralizing antibody and T cell responses in mice and nonhuman primates , 2020, Science Translational Medicine.

[66]  Ishtiaque Ahammad,et al.  Designing a novel mRNA vaccine against SARS-CoV-2: An immunoinformatics approach , 2020, International Journal of Biological Macromolecules.

[67]  Somdeb Mitra,et al.  5′-UTR recruitment of the translation initiation factor eIF4GI or DAP5 drives cap-independent translation of a subset of human mRNAs , 2020, The Journal of Biological Chemistry.

[68]  D. Fuller,et al.  Amplifying RNA Vaccine Development. , 2020, The New England journal of medicine.

[69]  O. Rossbach,et al.  Production and Purification of Artificial Circular RNA Sponges for Application in Molecular Biology and Medicine , 2020, Methods and protocols.

[70]  W. Gilbert,et al.  Direct analysis of ribosome targeting illuminates thousand-fold regulation of translation initiation , 2020, bioRxiv.

[71]  D. Weissman,et al.  Recent advances in mRNA vaccine technology. , 2020, Current opinion in immunology.

[72]  M. Komiyama,et al.  Preferential production of RNA rings by T4 RNA ligase 2 without any splint through rational design of precursor strand , 2020, Nucleic acids research.

[73]  M. Ringenberg,et al.  Nonclinical safety assessment of repeated administration and biodistribution of a novel rabies self-amplifying mRNA vaccine in rats. , 2020, Regulatory toxicology and pharmacology : RTP.

[74]  N. Barron,et al.  Reinventing the Wheel: Synthetic Circular RNAs for Mammalian Cell Engineering. , 2020, Trends in biotechnology.

[75]  S. Raj,et al.  Exploring the Zika Genome to Design a Potential Multiepitope Vaccine Using an Immunoinformatics Approach , 2020, International Journal of Peptide Research and Therapeutics.

[76]  Ö. Türeci,et al.  A Trans-amplifying RNA Vaccine Strategy for Induction of Potent Protective Immunity. , 2020, Molecular therapy : the journal of the American Society of Gene Therapy.

[77]  C. Lacroix,et al.  Tailoring mRNA Vaccine to Balance Innate/Adaptive Immune Response. , 2019, Trends in molecular medicine.

[78]  Howard Y. Chang,et al.  N6-Methyladenosine Modification Controls Circular RNA Immunity. , 2019, Molecular cell.

[79]  C. Chiang,et al.  Endogenous Nucleic Acid Recognition by RIG-I-Like Receptors and cGAS. , 2019, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[80]  B. Baradaran,et al.  Liposome and immune system interplay: Challenges and potentials. , 2019, Journal of controlled release : official journal of the Controlled Release Society.

[81]  I. Poser,et al.  Defective ribosomal products challenge nuclear function by impairing nuclear condensate dynamics and immobilizing ubiquitin , 2019, The EMBO journal.

[82]  Mei-Sheng Xiao,et al.  An improved method for circular RNA purification using RNase R that efficiently removes linear RNAs containing G-quadruplexes or structured 3′ ends , 2019, Nucleic acids research.

[83]  S. Müller,et al.  Engineering of hairpin ribozyme variants for RNA recombination and splicing , 2019, Annals of the New York Academy of Sciences.

[84]  Tyler E. Wagner,et al.  In vitro evolution of enhanced RNA replicons for immunotherapy , 2019, Scientific Reports.

[85]  Daniel G. Anderson,et al.  RNA Circularization Diminishes Immunogenicity and Can Extend Translation Duration In Vivo. , 2019, Molecular cell.

[86]  Qi Zhang,et al.  Translation of the circular RNA circβ-catenin promotes liver cancer cell growth through activation of the Wnt pathway , 2019, Genome Biology.

[87]  J. Ulmer,et al.  mRNA as a Transformative Technology for Vaccine Development to Control Infectious Diseases. , 2019, Molecular therapy : the journal of the American Society of Gene Therapy.

[88]  P. Mason,et al.  Self-Amplifying RNA Vaccines for Venezuelan Equine Encephalitis Virus Induce Robust Protective Immunogenicity in Mice. , 2019, Molecular therapy : the journal of the American Society of Gene Therapy.

[89]  Ö. Türeci,et al.  Improving mRNA-Based Therapeutic Gene Delivery by Expression-Augmenting 3' UTRs Identified by Cellular Library Screening. , 2019, Molecular therapy : the journal of the American Society of Gene Therapy.

[90]  U. Şahin,et al.  A Facile Method for the Removal of dsRNA Contaminant from In Vitro-Transcribed mRNA , 2019, Molecular therapy. Nucleic acids.

[91]  K. Burke,et al.  Durable anticancer immunity from intratumoral administration of IL-23, IL-36γ, and OX40L mRNAs , 2019, Science Translational Medicine.

[92]  M. Helm,et al.  RNA Modifications Modulate Activation of Innate Toll-Like Receptors , 2019, Genes.

[93]  H. Youn,et al.  Development of an RNA Expression Platform Controlled by Viral Internal Ribosome Entry Sites. , 2019, Journal of microbiology and biotechnology.

[94]  Ellese Marie Carmona,et al.  Circular RNA: Design Criteria for Optimal Therapeutical Utility , 2019 .

[95]  C. Rudolph,et al.  Segmented poly(A) tails significantly reduce recombination of plasmid DNA without affecting mRNA translation efficiency or half-life , 2019, RNA.

[96]  C. Rudolph,et al.  Maximizing the Translational Yield of mRNA Therapeutics by Minimizing 5'-UTRs. , 2019, Tissue engineering. Part A.

[97]  A. Fisher,et al.  Structural basis for eukaryotic mRNA modification. , 2018, Current opinion in structural biology.

[98]  S. Buus,et al.  Efficient Induction of T Cells against Conserved HIV-1 Regions by Mosaic Vaccines Delivered as Self-Amplifying mRNA , 2018, Molecular therapy. Methods & clinical development.

[99]  C. Mayr.  What Are 3' UTRs Doing? , 2018, Cold Spring Harbor perspectives in biology.

[100]  Florian Krammer,et al.  Nucleoside-modified mRNA immunization elicits influenza virus hemagglutinin stalk-specific antibodies , 2018, Nature Communications.

[101]  P. McKay,et al.  Structural Components for Amplification of Positive and Negative Strand VEEV Splitzicons , 2018, Front. Mol. Biosci..

[102]  Daniel G. Anderson,et al.  Engineering circular RNA for potent and stable translation in eukaryotic cells , 2018, Nature Communications.

[103]  Anton P. McCaffrey,et al.  Uridine Depletion and Chemical Modification Increase Cas9 mRNA Activity and Reduce Immunogenicity without HPLC Purification , 2018, Molecular therapy. Nucleic acids.

[104]  Georg Seelig,et al.  Human 5′ UTR design and variant effect prediction from a massively parallel translation assay , 2018, Nature Biotechnology.

[105]  G. Mcinerney,et al.  The Enigmatic Alphavirus Non-Structural Protein 3 (nsP3) Revealing Its Secrets at Last , 2018, Viruses.

[106]  H. Lee,et al.  Emergence of synthetic mRNA: In vitro synthesis of mRNA and its applications in regenerative medicine. , 2018, Biomaterials.

[107]  L. Ng,et al.  Nonstructural Proteins of Alphavirus—Potential Targets for Drug Development , 2018, Viruses.

[108]  T. Morrison,et al.  Innate immune control of alphavirus infection. , 2018, Current opinion in virology.

[109]  Sadeem Ahmad,et al.  An origin of the immunogenicity of in vitro transcribed RNA , 2018, bioRxiv.

[110]  M. Koike,et al.  Optimization of mNeonGreen for Homo sapiens increases its fluorescent intensity in mammalian cells , 2018, PloS one.

[111]  D. Weissman,et al.  mRNA vaccines — a new era in vaccinology , 2018, Nature Reviews Drug Discovery.

[112]  Rhiju Das,et al.  Functional 5′ UTR mRNA structures in eukaryotic translation regulation and how to find them , 2017, Nature Reviews Molecular Cell Biology.

[113]  Gene W. Yeo,et al.  Short Poly(A) Tails are a Conserved Feature of Highly Expressed Genes , 2017, Nature Structural & Molecular Biology.

[114]  Tomasz K. Wirecki,et al.  MODOMICS: a database of RNA modification pathways. 2017 update , 2017, Nucleic Acids Res..

[115]  A. Mishra,et al.  Exploring dengue genome to construct a multi-epitope based subunit vaccine by utilizing immunoinformatics approach to battle against dengue infection , 2017, Scientific Reports.

[116]  Bettina Appel,et al.  In vitro circularization of RNA , 2017, RNA biology.

[117]  Howard Y. Chang,et al.  Sensing Self and Foreign Circular RNAs by Intron Identity. , 2017, Molecular cell.

[118]  J. Utikal,et al.  Personalized RNA mutanome vaccines mobilize poly-specific therapeutic immunity against cancer , 2017, Nature.

[119]  N. Rajewsky,et al.  Translation of CircRNAs , 2017, Molecular cell.

[120]  Pengpeng Xia,et al.  RNA sensors of the innate immune system and their detection of pathogens , 2017, IUBMB life.

[121]  Yang Zhang,et al.  Extensive translation of circular RNAs driven by N6-methyladenosine , 2017, Cell Research.

[122]  D. Weissman,et al.  Zika virus protection by a single low dose nucleoside modified mRNA vaccination , 2017, Nature.

[123]  Chengqi Yi,et al.  Chemical Modifications to RNA: A New Layer of Gene Expression Regulation. , 2017, ACS chemical biology.

[124]  Olivier Elemento,et al.  Reversible methylation of m6Am in the 5′ cap controls mRNA stability , 2016, Nature.

[125]  J. Marcotrigiano,et al.  RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling , 2016, mBio.

[126]  Daniel G. Anderson,et al.  Dendrimer-RNA nanoparticles generate protective immunity against lethal Ebola, H1N1 influenza, and Toxoplasma gondii challenges with a single dose , 2016, Proceedings of the National Academy of Sciences.

[127]  G. B. Robb,et al.  mRNA capping: biological functions and applications , 2016, Nucleic acids research.

[128]  A. Hinnebusch,et al.  Translational control by 5′-untranslated regions of eukaryotic mRNAs , 2016, Science.

[129]  Yoosik Kim,et al.  Regulation of Poly(A) Tail and Translation during the Somatic Cell Cycle. , 2016, Molecular cell.

[130]  V. Hornung,et al.  Recognition of Endogenous Nucleic Acids by the Innate Immune System. , 2016, Immunity.

[131]  A. Scharenberg,et al.  pEVL: A Linear Plasmid for Generating mRNA IVT Templates With Extended Encoded Poly(A) Sequences , 2016, Molecular therapy. Nucleic acids.

[132]  T. Rades,et al.  Liposome-Based Adjuvants for Subunit Vaccines: Formulation Strategies for Subunit Antigens and Immunostimulators , 2016, Pharmaceutics.

[133]  Z. Yakhini,et al.  Systematic discovery of cap-independent translation sequences in human and viral genomes , 2016, Science.

[134]  M. Cobb.  Who discovered messenger RNA? , 2015, Current Biology.

[135]  N. Sonenberg,et al.  Protein Synthesis Initiation in Eukaryotes: IRES‐mediated Internal Initiation , 2015 .

[136]  S. Müller,et al.  RNA circularization strategies in vivo and in vitro , 2015, Nucleic acids research.

[137]  Yang Wang,et al.  Efficient backsplicing produces translatable circular mRNAs , 2015, RNA.

[138]  D. Weissman.  mRNA transcript therapy , 2015, Expert review of vaccines.

[139]  G. Rodrigo,et al.  Engineering a Circular Riboregulator in Escherichia coli , 2014, bioRxiv.

[140]  Özlem Türeci,et al.  mRNA-based therapeutics — developing a new class of drugs , 2014, Nature Reviews Drug Discovery.

[141]  M. Joshi,et al.  Peptide Vaccine: Progress and Challenges , 2014, Vaccines.

[142]  Martin A. M. Reijns,et al.  RNA:DNA hybrids are a novel molecular pattern sensed by TLR9 , 2014, The EMBO journal.

[143]  D. Rio.  Expression and purification of active recombinant T7 RNA polymerase from E. coli. , 2013, Cold Spring Harbor protocols.

[144]  Wei-Chiang Shen,et al.  Fusion protein linkers: property, design and functionality. , 2013, Advanced drug delivery reviews.

[145]  S. Müller,et al.  RNA self‐processing: Formation of cyclic species and concatemers from a small engineered RNA , 2013, FEBS letters.

[146]  Sebastian D. Mackowiak,et al.  Circular RNAs are a large class of animal RNAs with regulatory potency , 2013, Nature.

[147]  Michael K. Slevin,et al.  Circular RNAs are abundant, conserved, and associated with ALU repeats. , 2013, RNA.

[148]  T. Schlake,et al.  Developing mRNA-vaccine technologies , 2012, RNA biology.

[149]  S. Weaver,et al.  Vector-Borne Transmission Imposes a Severe Bottleneck on an RNA Virus Population , 2012, PLoS pathogens.

[150]  O. Elemento,et al.  Comprehensive Analysis of mRNA Methylation Reveals Enrichment in 3′ UTRs and near Stop Codons , 2012, Cell.

[151]  M. Kupiec,et al.  Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq , 2012, Nature.

[152]  M. Brinton,et al.  Activation of Oas1a gene expression by type I IFN requires both STAT1 and STAT2 while only STAT2 is required for Oas1b activation. , 2012, Virology.

[153]  D. Weissman,et al.  Generating the optimal mRNA for therapy: HPLC purification eliminates immune activation and improves translation of nucleoside-modified, protein-encoding mRNA , 2011, Nucleic acids research.

[154]  A. Hinnebusch,et al.  Molecular Mechanism of Scanning and Start Codon Selection in Eukaryotes , 2011, Microbiology and Molecular Reviews.

[155]  A. Lambowitz,et al.  Group II introns: mobile ribozymes that invade DNA. , 2011, Cold Spring Harbor perspectives in biology.

[156]  Pirjo Spuul,et al.  Assembly of Alphavirus Replication Complexes from RNA and Protein Components in a Novel trans-Replication System in Mammalian Cells , 2011, Journal of Virology.

[157]  S. Plotkin,et al.  Vaccines: the Fourth Century , 2009, Clinical and Vaccine Immunology.

[158]  K. Kisich,et al.  Hairpin ribozyme-antisense RNA constructs can act as molecular lassos , 2008, Nucleic acids research.

[159]  S. Akira,et al.  Length-dependent recognition of double-stranded ribonucleic acids by retinoic acid–inducible gene-I and melanoma differentiation–associated gene 5 , 2008, The Journal of experimental medicine.

[160]  H. Čelešnik,et al.  The bacterial enzyme RppH triggers messenger RNA degradation by 5′ pyrophosphate removal , 2008, Nature.

[161]  H. Schild,et al.  Increased Antigen Presentation Efficiency by Coupling Antigens to MHC Class I Trafficking Signals1 , 2008, The Journal of Immunology.

[162]  H. Rammensee,et al.  Spontaneous cellular uptake of exogenous messenger RNA in vivo is nucleic acid-specific, saturable and ion dependent , 2007, Gene Therapy.

[163]  W. Dalton.  Faculty Opinions recommendation of A "silent" polymorphism in the MDR1 gene changes substrate specificity. , 2007 .

[164]  U. Şahin,et al.  Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. , 2006, Blood.

[165]  Gunther Hartmann,et al.  5'-Triphosphate RNA Is the Ligand for RIG-I , 2006, Science.

[166]  R. Bowater,et al.  Direct comparison of nick-joining activity of the nucleic acid ligases from bacteriophage T4. , 2006, The Biochemical journal.

[167]  Rino Rappuoli,et al.  Post‐genomic vaccine development , 2006, FEBS letters.

[168]  Marcin Feder,et al.  MODOMICS: a database of RNA modification pathways , 2005, Nucleic Acids Res..

[169]  D. Haussler,et al.  Evolutionarily conserved elements in vertebrate, insect, worm, and yeast genomes. , 2005, Genome research.

[170]  Houping Ni,et al.  Suppression of RNA recognition by Toll-like receptors: the impact of nucleoside modification and the evolutionary origin of RNA. , 2005, Immunity.

[171]  K. Lindblad-Toh,et al.  Systematic discovery of regulatory motifs in human promoters and 3′ UTRs by comparison of several mammals , 2005, Nature.

[172]  W. Cao,et al.  Deviation from major codons in the Toll‐like receptor genes is associated with low Toll‐like receptor expression , 2005, Immunology.

[173]  R. Rhoads,et al.  Novel "anti-reverse" cap analogs with superior translational properties. , 2003, RNA.

[174]  R. Rhoads,et al.  Synthesis and properties of mRNAs containing the novel "anti-reverse" cap analogs 7-methyl(3'-O-methyl)GpppG and 7-methyl (3'-deoxy)GpppG. , 2001, RNA.

[175]  H. L. Murray,et al.  Excision of group II introns as circles. , 2001, Molecular cell.

[176]  G. Kollias,et al.  Impaired on/off regulation of TNF biosynthesis in mice lacking TNF AU-rich elements: implications for joint and gut-associated immunopathologies. , 1999, Immunity.

[177]  M. Muckenthaler,et al.  Poly(A)-tail-promoted translation in yeast: implications for translational control. , 1998, RNA.

[178]  Eric T. Kool,et al.  Generation of circular RNAs and trans-cleaving catalytic RNAs by rolling transcription of circular DNA oligonucleotides encoding hairpin ribozymes , 1998, Nucleic Acids Res..

[179]  L. Wang,et al.  Oligoribonucleotide circularization by 'template-mediated' ligation with T4 RNA ligase: synthesis of circular hammerhead ribozymes. , 1998, Nucleic acids research.

[180]  K. Jarrell,et al.  Use of an engineered ribozyme to produce a circular human exon. , 1997, Nucleic acids research.

[181]  A. Suhrbier,et al.  Multi‐epitope DNA vaccines , 1997, Immunology and cell biology.

[182]  P. Sarnow,et al.  Initiation of protein synthesis by the eukaryotic translational apparatus on circular RNAs. , 1995, Science.

[183]  G. Rhodes,et al.  Self-replicating Semliki Forest virus RNA as recombinant vaccine. , 1994, Vaccine.

[184]  M. Ares,et al.  Synthesis of circular RNA in bacteria and yeast using RNA cyclase ribozymes derived from a group I intron of phage T4. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[185]  M. Puttaraju,et al.  Group I permuted intron-exon (PIE) sequences self-splice to produce circular exons. , 1992, Nucleic acids research.

[186]  Kathleen R. Cho,et al.  Scrambled exons , 1991, Cell.

[187]  M. Coca-Prados,et al.  Electron microscopic evidence for the circular form of RNA in the cytoplasm of eukaryotic cells , 1979, Nature.

[188]  D. Riesner,et al.  Viroids are single-stranded covalently closed circular RNA molecules existing as highly base-paired rod-like structures. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[189]  A. Kanampalliwar.  Reverse Vaccinology and Its Applications. , 2020, Methods in molecular biology.

[190]  Naoko Abe,et al.  Preparation of Circular RNA In Vitro. , 2018, Methods in molecular biology.

[191]  S. Müller,et al.  Synthesis and Engineering of Circular RNAs. , 2018, Methods in molecular biology.

[192]  Ph.D Breton Hornblower,et al.  Minding your caps and tails – considerations for functional mRNA synthesis , 2015 .

[193]  C. Mandl,et al.  Self-amplifying mRNA vaccines. , 2015, Advances in genetics.

[194]  D. Weissman,et al.  In vitro transcription of long RNA containing modified nucleosides. , 2013, Methods in molecular biology.

[195]  D. Weissman,et al.  HPLC purification of in vitro transcribed long RNA. , 2013, Methods in molecular biology.

[196]  R. O’Keefe,et al.  Splint ligation of RNA with T4 DNA ligase. , 2012, Methods in molecular biology.

[197]  M. Moore.  Joining RNA molecules with T4 DNA ligase. , 1999, Methods in molecular biology.

[198]  D. Melton,et al.  In vitro RNA synthesis with SP6 RNA polymerase. , 1987, Methods in enzymology.

[199]  O. Uhlenbeck,et al.  2 T4 RNA Ligase , 1982 .