Wolff, J. A. et al. Direct gene transfer into mouse muscle in vivo. Science 247, 1465–1468 (1990).
Jirikowski, G. F. et al. Reversal of diabetes insipidus in Brattleboro rats: intrahypothalamic injection of vasopressin mRNA. Science 255, 996–998 (1992).
Martinon, F. et al. Induction of virus-specific cytotoxic T lymphocytes in vivo by liposome-entrapped mRNA. Eur. J. Immunol. 23, 1719–1722 (1993).
Conry, R. M. et al. Characterization of a messenger RNA polynucleotide vaccine vector. Cancer Res. 55, 1397–1400 (1995).
Boczkowski, D. et al. Dendritic cells pulsed with RNA are potent antigen-presenting cells in vitro and in vivo. J. Exp. Med. 184, 465–472 (1996).
Qiu, P. et al. Gene gun delivery of mRNA in situ results in efficient transgene expression and genetic immunization. Gene Ther. 3, 262–268 (1996).
Mandl, C. W. et al. In vitro-synthesized infectious RNA as an attenuated live vaccine in a flavivirus model. Nature Med. 4, 1438–1440 (1998).
Zhou, W. Z. et al. RNA melanoma vaccine: induction of antitumor immunity by human glycoprotein 100 mRNA immunization. Hum. Gene Ther. 10, 2719–2724 (1999).
Hoerr, I. et al. In vivo application of RNA leads to induction of specific cytotoxic T lymphocytes and antibodies. Eur. J. Immunol. 30, 1–7 (2000).
Koido, S. et al. Induction of antitumor immunity by vaccination of dendritic cells transfected with MUC1 RNA. J. Immunol. 165, 5713–5719 (2000).
Schirmacher, V. et al. Intra-pinna anti-tumor vaccination with self-replicating infectious RNA or with DNA encoding a model tumor antigen and a cytokine. Gene Ther. 7, 1137–1147 (2000).
Heiser, A. et al. Autologous dendritic cells transfected with prostate-specific antigen RNA stimulate CTL responses against metastatic prostate tumors. J. Clin. Invest. 109, 409–417 (2002).
Morse, M. A. et al. The feasibility and safety of immunotherapy with dendritic cells loaded with CEA mRNA following neoadjuvant chemoradiotherapy and resection of pancreatic cancer. Int. J. Gastrointest Cancer 32, 1–6 (2002).
Morse, M. A. et al. Immunotherapy with autologous, human dendritic cells transfected with carcinoembryonic antigen mRNA. Cancer Invest. 21, 341–349 (2003).
Su, Z. et al. Telomerase mRNA-transfected dendritic cells stimulate antigen-specific CD8+ and CD4+ T cell responses in patients with metastatic prostate cancer. J. Immunol. 174, 3798–3807 (2005).
Weide, B. et al. Direct injection of protamine-protected mRNA: results of a phase 1/2 vaccination trial in metastatic melanoma patients. J. Immunother. 32, 498–507 (2009).
Rittig, S. M. et al. Intradermal vaccinations with RNA coding for TAA generate CD8+ and CD4+ immune responses and induce clinical benefit in vaccinated patients. Mol. Ther. 19, 990–999 (2011).
Wilgenhof, S. et al. A phase IB study on intravenous synthetic mRNA electroporated dendritic cell immunotherapy in pretreated advanced melanoma patients. Ann. Oncol. 24, 2686–2693 (2013).
Okumura, K. et al. Bax mRNA therapy using cationic liposomes for human malignant melanoma. J. Gene Med. 10, 910–917 (2008).
Mitchell, D. A. et al. Selective modification of antigen-specific T cells by RNA electroporation. Hum. Gene Ther. 19, 511–521 (2008).
Wang, Y. et al. Systemic delivery of modified mRNA encoding herpes simplex virus 1 thymidine kinase for targeted cancer gene therapy. Mol. Ther. 21, 358–367 (2013).
Zimmermann, O. et al. Successful use of mRNA-nucleofection for overexpression of interleukin-10 in murine monocytes/macrophages for anti-inflammatory therapy in a murine model of autoimmune myocarditis. J. Am. Heart Assoc. 1, e003293 (2012).
Zangi, L. et al. Modified mRNA directs the fate of heart progenitor cells and induces vascular regeneration after myocardial infarction. Nature Biotech. 31, 898–907 (2013).
Creusot, R. J. et al. A short pulse of IL-4 delivered by DCs electroporated with modified mRNA can both prevent and treat autoimmune diabetes in NOD mice. Mol. Ther. 18, 2112–2120 (2010).
Kormann, M. S. et al. Expression of therapeutic proteins after delivery of chemically modified mRNA in mice. Nature Biotech. 29, 154–157 (2011).
Kariko, K. et al. Increased erythropoiesis in mice injected with submicrogram quantities of pseudouridine-containing mRNA encoding erythropoietin. Mol. Ther. 20, 948–953 (2012).
Mays, L. E. et al. Modified Foxp3 mRNA protects against asthma through an IL-10-dependent mechanism. J. Clin. Invest. 123, 1216–1228 (2013).
Levy, O. et al. mRNA-engineered mesenchymal stem cells for targeted delivery of interleukin-10 to sites of inflammation. Blood 122, e23–e32 (2013).
Holtkamp, S. et al. Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. Blood 108, 4009–4017 (2006).
Karikó, K., Kuo, A. & Barnathan, E. Overexpression of urokinase receptor in mammalian cells following administration of the in vitro transcribed encoding mRNA. Gene Ther. 6, 1092–1100 (1999).
Kallen, K.-J. & Theß, A. A development that may evolve into a revolution in medicine: mRNA as the basis for novel, nucleotide-based vaccines and drugs. Ther. Adv. Vaccines 2, 10–31 (2014).
Li, Y. & Kiledjian, M. Regulation of mRNA decapping. Wiley Interdiscip. Rev. RNA 1, 253–265 (2010).
Martin, S. A., Paoletti, E. & Moss, B. Purification of mRNA guanylyltransferase and mRNA (guanine-7-) methyltransferase from vaccinia virions. J. Biol. Chem. 250, 9322–9329 (1975).
Malone, R. W., Felgner, P. L. & Verma, I. M. Cationic liposome-mediated RNA transfection. Proc. Natl Acad. Sci. USA 86, 6077–6081 (1989).
Stepinski, J. 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. RNA 7, 1486–1495 (2001).
Jemielity, J. et al. Novel “anti-reverse” cap analogs with superior translational properties. RNA 9, 1108–1122 (2003).
Mockey, M. et al. mRNA transfection of dendritic cells: synergistic effect of ARCA mRNA capping with poly(A) chains in cis and in trans for a high protein expression level. Biochem. Biophys. Res. Commun. 340, 1062–1068 (2006).
Rabinovich, P. M. et al. Synthetic messenger RNA as a tool for gene therapy. Hum. Gene Ther. 17, 1027–1035 (2006).
Grudzien-Nogalska, E. et al. Phosphorothioate cap analogs stabilize mRNA and increase translational efficiency in mammalian cells. RNA 13, 1745–1755 (2007).
Kowalska, J. et al. Synthesis and characterization of mRNA cap analogs containing phosphorothioate substitutions that bind tightly to eIF4E and are resistant to the decapping pyrophosphatase DcpS. RNA 14, 1119–1131 (2008).
Kuhn, A. N. et al. Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo. Gene Ther. 17, 961–971 (2010).
Gallie, D. R. The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev. 5, 2108–2116 (1991).
Korner, C. G. & Wahle, E. Poly(A) tail shortening by a mammalian poly(A)-specific 3′-exoribonuclease. J. Biol. Chem. 272, 10448–10456 (1997).
Martin, G. & Keller, W. Tailing and 3′-end labeling of RNA with yeast poly(A) polymerase and various nucleotides. RNA 4, 226–230 (1998).
Ross, J. & Sullivan, T. Half-lives of β and γ globin messenger RNAs and of protein synthetic capacity in cultured human reticulocytes. Blood 66, 1149–1154 (1985).
Zinckgraf, J. W. & Silbart, L. K. Modulating gene expression using DNA vaccines with different 3′-UTRs influences antibody titer, seroconversion and cytokine profiles. Vaccine 21, 1640–1649 (2003).
Bergman, N. et al. Lsm proteins bind and stabilize RNAs containing 5′ poly(A) tracts. Nature Struct. Mol. Biol. 14, 824–831 (2007).
Kuhn, A. N. et al. mRNA as a versatile tool for exogenous protein expression. Curr. Gene Ther. 12, 347–361 (2012).
Chen, C. Y. & Shyu, A. B. AU-rich elements: characterization and importance in mRNA degradation. Trends Biochem. Sci. 20, 465–470 (1995).
Gustafsson, C., Govindarajan, S. & Minshull, J. Codon bias and heterologous protein expression. Trends Biotechnol. 22, 346–353 (2004).
Cannarozzi, G. et al. A role for codon order in translation dynamics. Cell 141, 355–367 (2010).
Bossi, L. & Ruth, J. R. The influence of codon context on genetic code translation. Nature 286, 123–127 (1980).
Gustafsson, C. et al. Engineering genes for predictable protein expression. Protein Expr. Purif. 83, 37–46 (2012).
Van Gulck, E. R. A. et al. Efficient stimulation of HIV-1-specific T cells using dendritic cells electroporated with mRNA encoding autologous HIV-1 Gag and Env proteins. Blood 107, 1818–1827 (2006).
Kimchi-Sarfaty, C. et al. A “silent” polymorphism in the MDR1 gene changes substrate specificity. Science 315, 525–528 (2007).
Malarkannan, S. et al. Presentation of out-of-frame peptide/MHC class I complexes by a novel translation initiation mechanism. Immunity 10, 681–690 (1999).
Saulquin, X. et al. +1 frameshifting as a novel mechanism to generate a cryptic cytotoxic T lymphocyte epitope derived from human interleukin 10. J. Exp. Med. 195, 353–358 (2002).
Schwab, S. R. et al. Constitutive display of cryptic translation products by MHC class I molecules. Science 301, 1367–1371 (2003).
Bourquin, C. et al. Immunostimulatory RNA oligonucleotides trigger an antigen-specific cytotoxic T-cell and IgG2a response. Blood 109, 2953–2960 (2007).
Sander, L. E. et al. Detection of prokaryotic mRNA signifies microbial viability and promotes immunity. Nature 474, 385–389 (2011).
Weissman, D. et al. HIV gag mRNA transfection of dendritic cells (DC) delivers encoded antigen to MHC class I and II molecules, causes DC maturation, and induces a potent human in vitro primary immune response. J. Immunol. 165, 4710–4717 (2000).
Fotin-Mleczek, M. et al. Messenger RNA-based vaccines with dual activity induce balanced TLR-7 dependent adaptive immune responses and provide antitumor activity. J. Immunother. 34, 1–15 (2011).
Karikó, K. et al. Generating the optimal mRNA for therapy: HPLC purification eliminates immune activation and improves translation of nucleoside-modified, protein-encoding mRNA. Nucleic Acids Res. 39, e142 (2011).
Rettig, L. et al. Particle size and activation threshold: a new dimension of danger signaling. Blood 115, 4533–4541 (2010).
Alexopoulou, L. et al. Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3. Nature 413, 732–738 (2001).
Diebold, S. S. et al. Innate antiviral responses by means of TLR7-mediated recognition of single-stranded RNA. Science 303, 1529–1531 (2004).
Heil, F. et al. Species-specific recognition of single-stranded RNA via Toll-like receptor 7 and 8. Science 303, 1526–1529 (2004).
Diebold, S. S. et al. Nucleic acid agonists for Toll-like receptor 7 are defined by the presence of uridine ribonucleotides. Eur. J. Immunol. 36, 3256–3267 (2006).
Yoneyama, M. et al. The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nature Immunol. 5, 730–737 (2004).
Yoneyama, M. et al. Shared and unique functions of the DExD/H-Box helicases RIG-I, MDA5, and LGP2 in antiviral innate immunity. J. Immunol. 175, 2851–2858 (2005).
Schlee, M. et al. Recognition of 5′ triphosphate by RIG-I helicase requires short blunt double-stranded RNA as contained in panhandle of negative-strand virus. Immunity 31, 25–34 (2009).
Pichlmair, A. et al. Activation of MDA5 requires higher-order RNA structures generated during virus infection. J. Virol. 83, 10761–10769 (2009).
Zust, R. et al. Ribose 2′-O-methylation provides a molecular signature for the distinction of self and non-self mRNA dependent on the RNA sensor Mda5. Nature Immunol. 12, 137–143 (2011).
Balachandran, S. et al. Essential role for the dsRNA-dependent protein kinase PKR in innate immunity to viral infection. Immunity 13, 129–141 (2000).
Karikó, K. et al. Suppression of RNA recognition by Toll-like receptors: the impact of nucleoside modification and the evolutionary origin of RNA. Immunity 23, 165–175 (2005).
Hornung, V. et al. 5′-triphosphate RNA is the ligand for RIG-I. Science 314, 994–997 (2006).
Nallagatla, S. R. & Bevilacqua, P. C. Nucleoside modifications modulate activation of the protein kinase PKR in an RNA structure-specific manner. RNA 14, 1201–1213 (2008).
Anderson, B. R. et al. Incorporation of pseudouridine into mRNA enhances translation by diminishing PKR activation. Nucleic Acids Res. 38, 5884–5892 (2010).
Lorenz, C. et al. Protein expression from exogenous mRNA: uptake by receptor-mediated endocytosis and trafficking via the lysosomal pathway. RNA Biol. 8, 627–636 (2011).
Valadi, H. et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nature Cell Biol. 9, 654–659 (2007).
Diken, M. et al. Selective uptake of naked vaccine RNA by dendritic cells is driven by macropinocytosis and abrogated upon DC maturation. Gene Ther. 18, 702–708 (2011).
Wang, W. et al. Non-viral gene delivery methods. Curr. Pharm. Biotechnol. 14, 46–60 (2013).
Neumann, E. et al. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1, 841–845 (1982).
Van Tendeloo, V. F. et al. Nonviral transfection of distinct types of human dendritic cells: high-efficiency gene transfer by electroporation into hematopoietic progenitor- but not monocyte-derived dendritic cells. Gene Ther. 5, 700–707 (1998).
Kyte, J. A. et al. Phase I/II trial of melanoma therapy with dendritic cells transfected with autologous tumor-mRNA. Cancer Gene Ther. 13, 905–918 (2006).
Van Driessche, A. et al. Clinical-grade manufacturing of autologous mature mRNA-electroporated dendritic cells and safety testing in acute myeloid leukemia patients in a phase I dose-escalation clinical trial. Cytotherapy 11, 653–668 (2009).
Van Tendeloo, V. F. et al. Induction of complete and molecular remissions in acute myeloid leukemia by Wilms' tumor 1 antigen-targeted dendritic cell vaccination. Proc. Natl Acad. Sci. USA 107, 13824–13829 (2010).
Wilgenhof, S. et al. Therapeutic vaccination with an autologous mRNA electroporated dendritic cell vaccine in patients with advanced melanoma. J. Immunother. 34, 448–456 (2011).
Van Nuffel, A. M. et al. Intravenous and intradermal TriMix-dendritic cell therapy results in a broad T-cell response and durable tumor response in a chemorefractory stage IV-M1c melanoma patient. Cancer Immunol. Immunother. 61, 1033–1043 (2012).
Van Nuffel, A. M. et al. Dendritic cells loaded with mRNA encoding full-length tumor antigens prime CD4+ and CD8+ T cells in melanoma patients. Mol. Ther. 20, 1063–1074 (2012).
Geng, T. et al. Transfection of cells using flow-through electroporation based on constant voltage. Nature Protoc. 6, 1192–1208 (2011).
Weide, B. et al. Results of the first phase I/II clinical vaccination trial with direct injection of mRNA. J. Immunother. 31, 180–188 (2008).
Wang, T., Upponi, J. R. & Torchilin, V. P. Design of multifunctional non-viral gene vectors to overcome physiological barriers: dilemmas and strategies. Int. J. Pharm. 427, 3–20 (2012).
Semple, S. C. et al. Rational design of cationic lipids for siRNA delivery. Nature Biotech. 28, 172–176 (2010).
Coelho, T. et al. Safety and efficacy of RNAi therapy for transthyretin amyloidosis. N. Engl. J. Med. 369, 819–829 (2013).
Granstein, R. D., Ding, W. & Ozawa, H. Induction of anti-tumor immunity with epidermal cells pulsed with tumor-derived RNA or intradermal administration of RNA. J. Invest. Dermatol. 114, 632–636 (2000).
Kreiter, S. et al. Intranodal vaccination with naked antigen-encoding RNA elicits potent prophylactic and therapeutic antitumoral immunity. Cancer Res. 70, 9031–9040 (2010).
Kreiter, S. et al. FLT3 ligand enhances the cancer therapeutic potency of naked RNA vaccines. Cancer Res. 71, 6132–6142 (2011).
Van Lint, S. et al. Preclinical evaluation of TriMix and antigen mRNA-based antitumor therapy. Cancer Res. 72, 1661–1671 (2012).
Petsch, B. et al. Protective efficacy of in vitro synthesized, specific mRNA vaccines against influenza A virus infection. Nature Biotech. 30, 1210–1216 (2012).
Geall, A. J. et al. Nonviral delivery of self-amplifying RNA vaccines. Proc. Natl Acad. Sci. USA 109, 14604–14609 (2012).
Uchida, S. et al. In vivo messenger RNA introduction into the central nervous system using polyplex nanomicelle. PLoS ONE 8, e56220 (2013).
Azarmi, S., Roa, W. H. & Lobenberg, R. Targeted delivery of nanoparticles for the treatment of lung diseases. Adv. Drug Deliv. Rev. 60, 863–875 (2008).
Torchilin, V. Tumor delivery of macromolecular drugs based on the EPR effect. Adv. Drug Deliv. Rev. 63, 131–135 (2011).
van der Bruggen, P. et al. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science 254, 1643–1647 (1991).
Sahin, U. et al. Human neoplasms elicit multiple specific immune responses in the autologous host. Proc. Natl Acad. Sci. USA 92, 11810–11813 (1995).
Nair, S. K. et al. Induction of cytotoxic T cell responses and tumor immunity against unrelated tumors using telomerase reverse transcriptase RNA transfected dendritic cells. Nature Med. 6, 1011–1017 (2000).
Nair, S. K. et al. Induction of primary carcinoembryonic antigen (CEA)-specific cytotoxic T lymphocytes in vitro using human dendritic cells transfected with RNA. Nature Biotech. 16, 364–369 (1998).
Morse, M. A. et al. Optimization of the sequence of antigen loading and CD40-ligand-induced maturation of dendritic cells. Cancer Res. 58, 2965–2968 (1998).
Van Lint, S. et al. mRNA: From a chemical blueprint for protein production to an off-the-shelf therapeutic. Hum. Vaccin. Immunother. 9, 265–274 (2013).
Kreiter, S. et al. Tumor vaccination using messenger RNA: prospects of a future therapy. Curr. Opin. Immunol. 23, 399–406 (2011).
Cisco, R. M. et al. Induction of human dendritic cell maturation using transfection with RNA encoding a dominant positive Toll-like receptor 4. J. Immunol. 172, 7162–7168 (2004).
Bonehill, A. et al. Enhancing the T-cell stimulatory capacity of human dendritic cells by co-electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA. Mol. Ther. 16, 1170–1180 (2008).
Calderhead, D. M. et al. Cytokine maturation followed by CD40L mRNA electroporation results in a clinically relevant dendritic cell product capable of inducing a potent proinflammatory CTL response. J. Immunother. 31, 731–741 (2008).
Routy, J.-P. et al. Immunologic activity and safety of autologous HIV RNA-electroporated dendritic cells in HIV-1 infected patients receiving antiretroviral therapy. Clin. Immunol. 134, 140–147 (2010).
Bontkes, H. J. et al. Dendritic cells transfected with interleukin-12 and tumor-associated antigen messenger RNA induce high avidity cytotoxic T cells. Gene Ther. 14, 366–375 (2007).
Aarntzen, E. H. J. G. et al. Vaccination with mRNA-electroporated dendritic cells induces robust tumor antigen-specific CD4+ and CD8+ T cells responses in stage III and IV melanoma patients. Clin. Cancer Res. 18, 5460–5470 (2012).
Pascolo, S. Vaccination with messenger RNA (mRNA). Handb Exp. Pharmacol. 183, 221–235 (2008).
Carralot, J. P. et al. Polarization of immunity induced by direct injection of naked sequence-stabilized mRNA vaccines. Cell. Mol. Life Sci. 61, 2418–2424 (2004).
Scheel, B. et al. Toll-like receptor-dependent activation of several human blood cell types by protamine-condensed mRNA. Eur. J. Immunol. 35, 1557–1566 (2005).
Fotin-Mleczek, M. et al. Highly potent mRNA based cancer vaccines represent an attractive platform for combination therapies supporting an improved therapeutic effect. J. Gene Med. 14, 428–439 (2012).
Kreiter, S. et al. Simultaneous ex vivo quantification of antigen-specific CD4+ and CD8+ T cell responses using in vitro transcribed RNA. Cancer Immunol. Immunother. 56, 1577–1587 (2007).
Kreiter, S. et al. Increased antigen presentation efficiency by coupling antigens to MHC class I trafficking signals. J. Immunol. 180, 309–318 (2008).
Diken, M. et al. Antitumor vaccination with synthetic mRNA: strategies for in vitro and in vivo preclinical studies. Methods Mol. Biol. 969, 235–246 (2013).
Diken, M. et al. mTOR inhibition improves antitumor effects of vaccination with antigen-encoding RNA. Cancer Immunol. Res. 1, 386–392 (2013).
Castle, J. C. et al. Exploiting the mutanome for tumor vaccination. Cancer Res. 72, 1081–1091 (2012).
Kreiter, S. et al. Targeting the tumor mutanome for personalized vaccination therapy. Oncoimmunology 1, 768–769 (2012).
Britten, C. M. et al. The regulatory landscape for actively personalized cancer immunotherapies. Nature Biotech. 31, 880–882 (2013).
Zhao, Y. et al. Multiple injections of electroporated autologous T cells expressing a chimeric antigen receptor mediate regression of human disseminated tumor. Cancer Res. 70, 9053–9061 (2010).
Almasbak, H. et al. Transiently redirected T cells for adoptive transfer. Cytotherapy 13, 629–640 (2011).
Barrett, D. M. et al. Treatment of advanced leukemia in mice with mRNA engineered T cells. Hum. Gene Ther. 22, 1575–1586 (2011).
Barrett, D. M. et al. Regimen specific effects of RNA-modified chimeric antigen receptor T cells in mice with advanced leukemia. Hum. Gene Ther. 24, 717–727 (2013).
Hekele, A. et al. Rapidly produced SAM® vaccine against H7N9 influenza is immunogenic in mice. Emerg. Microbes Infect. 2, e52 (2013).
Allard, S. D. et al. A phase I/IIa immunotherapy trial of HIV-1-infected patients with Tat, Rev and Nef expressing dendritic cells followed by treatment interruption. Clin. Immunol. 142, 252–268 (2012).
Van Gulck, E. et al. mRNA-based dendritic cell vaccination induces potent antiviral T-cell responses in HIV-1-infected patients. AIDS 26, F1–F12 (2012).
Liljestrom, P. & Garoff, H. A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Biotechnology 9, 1356–1361 (1991).
Zhou, X. et al. Self-replicating Semliki Forest virus RNA as recombinant vaccine. Vaccine 12, 1510–1514 (1994).
Ulmer, J. B. et al. RNA-based vaccines. Vaccine 30, 4414–4418 (2012).
Geall, A. J., Mandl, C. W. & Ulmer, J. B. RNA: the new revolution in nucleic acid vaccines. Semin. Immunol. 25, 152–159 (2013).
Fleeton, M. N. et al. Self-replicative RNA vaccines elicit protection against influenza A virus, respiratory syncytial virus, and a tickborne encephalitis virus. J. Infect. Dis. 183, 1395–1398 (2001).
Anraku, I. et al. Kunjin virus replicon vaccine vectors induce protective CD8+ T-cell immunity. J. Virol. 76, 3791–3799 (2002).
Greer, C. E. et al. A chimeric alphavirus RNA replicon gene-based vaccine for human parainfluenza virus type 3 induces protective immunity against intranasal virus challenge. Vaccine 25, 481–489 (2007).
Valenta, R. et al. From allergen genes to allergy vaccines. Annu. Rev. Immunol. 28, 211–241 (2010).
Raz, E. et al. Preferential induction of a Th1 immune response and inhibition of specific IgE antibody formation by plasmid DNA immunization. Proc. Natl Acad. Sci. USA 93, 5141–5145 (1996).
Chua, K. Y., Kuo, I. C. & Huang, C. H. DNA vaccines for the prevention and treatment of allergy. Curr. Opin. Allergy Clin. Immunol. 9, 50–54 (2009).
Slater, J. E. et al. The latex allergen Hev b 5 transcript is widely distributed after subcutaneous injection in BALB/c mice of its DNA vaccine. J. Allergy Clin. Immunol. 102, 469–475 (1998).
Roesler, E. et al. Immunize and disappear-safety-optimized mRNA vaccination with a panel of 29 allergens. J. Allergy Clin. Immunol, 124, 1070–1077.e11 (2009).
Weiss, R. et al. Prophylactic mRNA vaccination against allergy. Curr. Opin. Allergy Clin. Immunol. 10, 567–574 (2010).
Kolarich, D. et al. Comprehensive glyco-proteomic analysis of human α1-antitrypsin and its charge isoforms. Proteomics 6, 3369–3380 (2006).
Seidah, N. G. & Chrétien, M. Proprotein and prohormone convertases: a family of subtilases generating diverse bioactive polypeptides. Brain Res. 848, 45–62 (1999).
Nakayama, K. Furin: a mammalian subtilisin/Kex2p-like endoprotease involved in processing of a wide variety of precursor proteins. Biochem. J. 327, 625–635 (1997).
Seidah, N. G. et al. Precursor convertases: an evolutionary ancient, cell-specific, combinatorial mechanism yielding diverse bioactive peptides and proteins. Ann. NY Acad. Sci. 839, 9–24 (1998).
Barash, S., Wang, W. & Shi, Y. Human secretory signal peptide description by hidden Markov model and generation of a strong artificial signal peptide for secreted protein expression. Biochem. Biophys. Res. Commun. 294, 835–842 (2002).
Fattori, E. et al. Gene electro-transfer of an improved erythropoietin plasmid in mice and non-human primates. J. Gene Med. 7, 228–236 (2005).
Roberts, A. A. et al. Engineering factor Viii for hemophilia gene therapy. J. Genet. Syndr. Gene Ther. 1, S1–006 (2011).
Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006).
Warren, L. et al. Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell 7, 618–630 (2010).
Warren, L. et al. Feeder-free derivation of human induced pluripotent stem cells with messenger RNA. Sci. Rep. 2, 657 (2012).
Mandal, P. K. & Rossi, D. J. Reprogramming human fibroblasts to pluripotency using modified mRNA. Nature Protoc. 8, 568–582 (2013).
Bernal, J. A. RNA-based tools for nuclear reprogramming and lineage-conversion: towards clinical applications. J. Cardiovasc. Transl. Res. 6, 956–968 (2013).
Miller, J. D. et al. Human iPSC-based modeling of late-onset disease via progerin-induced aging. Cell Stem Cell 13, 691–705 (2013).
Scott, C. W., Peters, M. F. & Dragan, Y. P. Human induced pluripotent stem cells and their use in drug discovery for toxicity testing. Toxicol. Lett. 219, 49–58 (2013).
Okano, H. et al. Steps toward safe cell therapy using induced pluripotent stem cells. Circ. Res. 112, 523–533 (2013).
Miller, J. C. et al. An improved zinc-finger nuclease architecture for highly specific genome editing. Nature Biotech. 25, 778–785 (2007).
Hockemeyer, D. et al. Genetic engineering of human pluripotent cells using TALE nucleases. Nature Biotech. 29, 731–734 (2011).
Mali, P. et al. RNA-guided human genome engineering via Cas9. Science 339, 823–826 (2013).
Fu, Y. et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotech. 31, 822–826 (2013).
Doyon, Y. et al. Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases. Nature Biotech. 26, 702–708 (2008).
Tesson, L. et al. Knockout rats generated by embryo microinjection of TALENs. Nature Biotech. 29, 695–696 (2011).
Wang, H. et al. One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153, 910–918 (2013).
Wefers, B. et al. Generation of targeted mouse mutants by embryo microinjection of TALEN mRNA. Nature Protocols 8, 2355–2379 (2013).
Geurts, A. M. et al. Knockout rats via embryo microinjection of zinc-finger nucleases. Science 325, 433 (2009).
Shen, B. et al. Generation of gene-modified mice via Cas9/RNA-mediated gene targeting. Cell Res. 23, 720–723 (2013).
Yang, D. et al. Effective gene targeting in rabbits using RNA-guided Cas9 nucleases. J. Mol. Cell Biol. 6, 97–99 (2014).
Ma, Y. et al. Generating rats with conditional alleles using CRISPR/Cas9. Cell Res. 24, 122–125 (2014).
Niu, Y. et al. Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos. Cell 156, 836–843 (2014).
Dupuy, A. J. et al. Mammalian germ-line transgenesis by transposition. Proc. Natl Acad. Sci. USA 99, 4495–4499 (2002).
Wilber, A. et al. RNA as a source of transposase for sleeping beauty-mediated gene insertion and expression in somatic cells and tissues. Mol. Ther. 13, 625–630 (2006).
Sumiyama, K., Kawakami, K. & Yagita, K. A simple and highly efficient transgenesis method in mice with the Tol2 transposon system and cytoplasmic microinjection. Genomics 95, 306–311 (2010).
Suster, M. L., Sumiyama, K. & Kawakami, K. Transposon-mediated BAC transgenesis in zebrafish and mice. BMC Genomics 10, 477 (2009).
Furushima, K. et al. Insertional mutagenesis by a hybrid piggyBac and sleeping beauty transposon in the rat. Genetics 192, 1235–1248 (2012).
Bire, S. et al. Exogenous mRNA delivery and bioavailability in gene transfer mediated by piggyBac transposition. BMC Biotechnol. 13, 75 (2013).
Isaacs, A., Cox, R. A. & Rotem, Z. Foreign nucleic acids as the stimulus to make interferon. Lancet 282, 113–116 (1963).
Tytell, A. A. et al. Inducers of interferon and host resistance. 3. Double-stranded RNA from reovirus type 3 virions (reo 3-RNA). Proc. Natl Acad. Sci. USA 58, 1719–1722 (1967).
Anderson, B. R. et al. Nucleoside modifications in RNA limit activation of 2′-5′-oligoadenylate synthetase and increase resistance to cleavage by RNase L. Nuc. Acids Res. 39, 9329–9338 (2011).
Pollard, C. et al. Type I IFN counteracts the induction of antigen-specific immune responses by lipid-based delivery of mRNA vaccines. Mol. Ther. 21, 251–259 (2013).
Hwang, S. H. et al. B cell TLR7 expression drives anti-RNA autoantibody production and exacerbates disease in systemic lupus erythematosus-prone mice. J. Immunol. 189, 5786–5796 (2012).
Lipes, B. D. & Keene, J. D. Autoimmune epitopes in messenger RNA. RNA 8, 762–771 (2002).
Murphy, K. (ed) in Janeway's Immunobiology 367–408 (Garland Science Publishing, 2011).
Worobec, A. & Rosenberg, A. S. A risk-based approach to immunogenicity concerns of therapeutic protein products, part 1: considering consequences of the immune response to a protein. BioPharm International 22–26 (2004).
Koren, E. et al. Recommendations on risk-based strategies for detection and characterization of antibodies against biotechnology products. J. Immunol. Methods 333, 1–9 (2008).
Casadevall, N. et al. Pure red-cell aplasia and antierythropoietin antibodies in patients treated with recombinant erythropoietin. N. Engl. J. Med. 346, 469–475 (2002).
Gao, G. et al. Erythropoietin gene therapy leads to autoimmune anemia in macaques. Blood 103, 3300–3302 (2004).
Kromminga, A. & Schellekens, H. Antibodies against erythropoietin and other protein-based therapeutics: an overview. Ann, NY Acad. Sci. 1050, 257–265 (2005).
Czech, M. P., Aouadi, M. & Tesz, G. J. RNAi-based therapeutic strategies for metabolic disease. Nature Rev. Endocrinol. 7, 473–484 (2011).
Racanelli, V. & Rehermann, B. The liver as an immunological organ. Hepatology 43 (Suppl. 1), S54–S62 (2006).
Dyer, K. D. & Rosenberg, H. F. The RNase a superfamily: generation of diversity and innate host defense. Mol. Divers. 13, 13 (2006).
McKenzie, R. et al. Hepatic failure and lactic acidosis due to fialuridine (FIAU), an investigational nucleoside analogue for chronic hepatitis B. N. Engl. J. Med. 333, 1099–1105 (1995).
Lewis, W. Defective mitochondrial DNA replication and NRTIs: pathophysiological implications in AIDS cardiomyopathy. Am. J. Physiol. Heart Circ. Physiol. 284, H1–H9 (2003).
Griffiths, M. et al. Cloning of a human nucleoside transporter implicated in the cellular uptake of adenosine and chemotherapeutic drugs. Nature Med. 3, 89–93 (1997).
Lewis, W. et al. Fialuridine and its metabolites inhibit DNA polymerase γ at sites of multiple adjacent analog incorporation, decrease mtDNA abundance, and cause mitochondrial structural defects in cultured hepatoblasts. Proc. Natl Acad. Sci. USA 93, 3592–3597 (1996).
Lai, Y., Tse, C.-M. & Unadkat, J. D. Mitochondrial expression of the human equilibrative nucleoside transporter 1 (hENT1) results in enhanced mitochondrial toxicity of antiviral drugs. J. Biol. Chem. 279, 4490–4497 (2004).
Lee, E.-W. et al. Identification of the mitochondrial targeting signal of the human equilibrative nucleoside transporter 1 (hENT1): implications for interspecies differences in mitochondrial toxicity of fialuridine. J. Biol. Chem. 281, 16700–16706 (2006).
Yoshioka, N. et al. Efficient generation of human iPSCs by a synthetic self-replicative RNA. Cell Stem Cell 13, 246–254 (2013).
Dormitzer, P. R. et al. Synthetic generation of influenza vaccine viruses for rapid response to pandemics. Sci. Transl. Med. 5, 185ra68 (2013).
Prieels, J.-P. et al. Mastering industrialization of cell therapy products. BioProcess Int. 10, S12–S15 (2012).
Dahm, R. Friedrich Miescher and the discovery of DNA. Dev. Biol. 278, 274–288 (2005).
Brenner, S., Jacob, F. & Meselson, M. An unstable intermediate carrying information from genes to ribosomes for protein synthesis. Nature 190, 576–581 (1961).
Racaniello, V. R. & Baltimore, D. Cloned poliovirus complementary DNA is infectious in mammalian cells. Science 214, 916–919 (1981).
Rice, C. M. et al. Production of infectious RNA transcripts from Sindbis virus cDNA clones: mapping of lethal mutations, rescue of a temperature-sensitive marker, and in vitro mutagenesis to generate defined mutants. J. Virol. 61, 3809–3819 (1987).
Etchison, D. & Ehrenfeld, E. Comparison of replication complexes synthesizing poliovirus RNA. Virology 111, 33–46 (1981).
Mizutani, S. & Colonno, R. J. In vitro synthesis of an infectious RNA from cDNA clones of human rhinovirus type 14. J. Virol. 56, 628–632 (1985).
van der Werf, S. et al. Synthesis of infectious poliovirus RNA by purified T7 RNA polymerase. Proc. Natl Acad. Sci. USA 83, 2330–2334 (1986).
Khromykh, A. A. & Westaway, E. G. Subgenomic replicons of the flavivirus Kunjin: construction and applications. J. Virol. 71, 1497–1505 (1997).
Perri, S. et al. An alphavirus replicon particle chimera derived from venezuelan equine encephalitis and sindbis viruses is a potent gene-based vaccine delivery vector. J. Virol. 77, 10394–10403 (2003).
Rolls, M. M. et al. Novel infectious particles generated by expression of the vesicular stomatitis virus glycoprotein from a self-replicating RNA. Cell 79, 497–506 (1994).
Xiong, C. et al. Sindbis virus: an efficient, broad host range vector for gene expression in animal cells. Science 243, 1188–1191 (1989).
Ying, H. et al. Cancer therapy using a self-replicating RNA vaccine. Nature Med. 5, 823–827 (1999).
Lundstrom, K. Alphaviruses in gene therapy. Viruses 1, 13–25 (2009).
Hewson, R. RNA viruses: emerging vectors for vaccination and gene therapy. Mol. Med. Today 6, 28–35 (2000).
Lundin, P. Is silence still golden? Mapping the RNAi patent landscape. Nature Biotech. 29, 493–497 (2011).
Modrak-Wojcik, A. et al. Eukaryotic translation initiation is controlled by cooperativity effects within ternary complexes of 4E-BP1, eIF4E, and the mRNA 5′ cap. FEBS Lett. 587, 3928–3934 (2013).
Rau, M. et al. A reevaluation of the cap-binding protein, eIF4E, as a rate-limiting factor for initiation of translation in reticulocyte lysate. J. Biol. Chem. 271, 8983–8990 (1996).
Wells, S. E. et al. Circularization of mRNA by eukaryotic translation initiation factors. Mol. Cell 2, 135–140 (1998).
Houseley, J. & Tollervey, D. The many pathways of RNA degradation. Cell 136, 763–776 (2009).
Balagopal, V., Fluch, L. & Nissan, T. Ways and means of eukaryotic mRNA decay. Biochim. Biophys. Acta 1819, 593–603 (2012).
Shyu, A. B., Wilkinson, M. F. & van Hoof, A. Messenger RNA regulation: to translate or to degrade. EMBO J. 27, 471–481 (2008).
Tomecki, R. & Dziembowski, A. Novel endoribonucleases as central players in various pathways of eukaryotic RNA metabolism. RNA 16, 1692–1724 (2010).
Li, W. M., Barnes, T. & Lee, C. H. Endoribonucleases — enzymes gaining spotlight in mRNA metabolism. FEBS J. 277, 627–641 (2010).
Wilusz, J. RNA stability: is it the endo' the world as we know it? Nature Struct. Mol. Biol. 16, 9–10 (2009).
Garneau, N. L., Wilusz, J. & Wilusz, C. J. The highways and byways of mRNA decay. Nature Rev. Mol. Cell Biol. 8, 113–126 (2007).
Bevan, M. J. Cross-priming. Nature Immunol. 7, 363–365 (2006).
Thomsen, L. B. et al. Nanoparticle-derived non-viral genetic transfection at the blood–brain barrier to enable neuronal growth factor delivery by secretion from brain endothelium. Curr. Med. Chem. 18, 3330–3334 (2011).
Hayashi, S. et al. Autocrine-paracrine effects of overexpression of hepatocyte growth factor gene on growth of endothelial cells. Biochem. Biophys. Res. Commun. 220, 539–545 (1996).
Zeis, M. et al. Generation of cytotoxic responses in mice and human individuals against hematological malignancies using survivin-RNA-transfected dendritic cells. J. Immunol. 170, 5391–5397 (2003).
Siegel, S. et al. Induction of cytotoxic T-cell responses against the oncofetal antigen-immature laminin receptor for the treatment of hematologic malignancies. Blood 102, 4416–4423 (2003).
Yoon, S. H. et al. Adoptive immunotherapy using human peripheral blood lymphocytes transferred with RNA encoding Her-2/neu-specific chimeric immune receptor in ovarian cancer xenograft model. Cancer Gene Ther. 16, 489–497 (2009).
Rabinovich, P. M. et al. Chimeric receptor mRNA transfection as a tool to generate antineoplastic lymphocytes. Hum. Gene Ther. 20, 51–61 (2009).
Bonehill, A. et al. Single-step antigen loading and activation of dendritic cells by mRNA electroporation for the purpose of therapeutic vaccination in melanoma patients. Clin. Cancer Res. 15, 3366–3375 (2009).
Maus, M. V. et al. T cells expressing chimeric antigen receptors can cause anaphylaxis in humans. Cancer Immunol. Res. 1, 26–31 (2013).
Benteyn, D. et al. Characterization of CD8+ T-cell responses in the peripheral blood and skin injection sites of melanoma patients treated with mRNA electroporated autologous dendritic cells (TriMixDC-MEL). Biomed Res Int http://dx.doi.org/10.1155/2013/976383 (2013).
Lorenzi, J. C. et al. Intranasal vaccination with messenger RNA as a new approach in gene therapy: use against tuberculosis. BMC Biotechnol. 10, 77 (2010).
Wood, A. J. et al. Targeted genome editing across species using ZFNs and TALENs. Science 333, 307 (2011).
Davies, B. et al. Site specific mutation of the Zic2 locus by microinjection of TALEN mRNA in mouse CD1, C3H and C57BL/6J oocytes. PLoS ONE 8, e60216 (2013).
Karikó, K. et al. Incorporation of pseudouridine into mRNA yields superior nonimmunogenic vector with increased translational capacity and biological stability. Mol Ther. 16, 1833–1840 (2008).
Angel, M. & Yanik, M. F. Innate immune suppression enables frequent transfection with RNA encoding reprogramming proteins. PLoS ONE 5, e11756 (2010).
Yakubov, E. et al. Reprogramming of human fibroblasts to pluripotent stem cells using mRNA of four transcription factors. Biochem. Biophys. Res. Commun. 394, 189–193 (2010).
Smull, C. E., Mallette, M. F. & Ludwig, E. H. The use of basic proteins to increase the infectivity of enterovirus ribonucleic acid. Biochem. Biophys. Res. Commun. 5, 247–249 (1961).
Papahadjopoulos, D. et al. Cochleate lipid cylinders: formation by fusion of unilamellar lipid vesicles. Biochim. Biophys. Acta 394, 483–491 (1975).
Dimitriadis, G. J. Translation of rabbit globin mRNA introduced by liposomes into mouse lymphocytes. Nature 274, 923–924 (1978).
Muthukrishnan, S., Both, G. W., Furuichi, Y. & Shatkin, A. J. 5′-Terminal 7-methylguanosine in eukaryotic mRNA is required for translation. Nature 255, 33–37 (1975).
Furuichi, Y. & Miura, K. A blocked structure at the 5′ terminus of mRNA from cytoplasmic polyhedrosis virus. Nature 253, 374–375 (1975).
Lockard, R. E. & Lingrel, J. B. The synthesis of mouse hemoglobin β-chains in a rabbit reticulocyte cell-free system programmed with mouse reticulocyte 9S RNA. Biochem. Biophys. Res. Commun. 37, 204–212 (1969).
Gurdon, J. B. et al. Use of frog eggs and oocytes for the study of messenger RNA and its translation in living cells. Nature. 233, 177–182 (1971).
Krieg, P. A. & Melton, D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucl. Acids Res. 12, 7057–7070 (1984).
Hwang, W. Y. et al. Efficient genome editing in zebrafish using a CRISPR-Cas system. Nature Biotech. 31, 227–229 (2013).