0000000000591061

AUTHOR

Tim Beissert

showing 5 related works from this author

Improving mRNA-Based Therapeutic Gene Delivery by Expression-Augmenting 3' UTRs Identified by Cellular Library Screening.

2019

Synthetic mRNA has emerged as a powerful tool for the transfer of genetic information, and it is being explored for a variety of therapeutic applications. Many of these applications require prolonged intracellular persistence of mRNA to improve bioavailability of the encoded protein. mRNA molecules are intrinsically unstable and their intracellular kinetics depend on the UTRs embracing the coding sequence, in particular the 3′ UTR elements. We describe here a novel and generally applicable cell-based selection process for the identification of 3′ UTRs that augment the expression of proteins encoded by synthetic mRNA. Moreover, we show, for two applications of mRNA therapeutics, namely, (1) …

Untranslated regionCellular differentiationRNA StabilityInduced Pluripotent Stem CellsBlood DonorsComputational biologyGene deliveryBiologyCancer Vaccines03 medical and health sciencesMice0302 clinical medicineDrug DiscoveryGeneticsCoding regionAnimalsHumansRNA MessengerInduced pluripotent stem cellMolecular BiologyGene3' Untranslated RegionsCells Cultured030304 developmental biologyGene LibraryPharmacology0303 health sciencesMessenger RNAMice Inbred BALB CVaccinationGene Transfer TechniquesGenetic TherapyFibroblastsCellular Reprogramming030220 oncology & carcinogenesisMolecular MedicineFemaleOriginal ArticleReprogrammingHalf-LifeMolecular therapy : the journal of the American Society of Gene Therapy
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Efficient Reprogramming of Human Fibroblasts and Blood-Derived Endothelial Progenitor Cells Using Nonmodified RNA for Reprogramming and Immune Evasion

2015

mRNA reprogramming results in the generation of genetically stable induced pluripotent stem (iPS) cells while avoiding the risks of genomic integration. Previously published mRNA reprogramming protocols have proven to be inconsistent and time-consuming and mainly restricted to fibroblasts, thereby demonstrating the need for a simple but reproducible protocol applicable to various cell types. So far there have been no published reports using mRNA to reprogram any cell type derived from human blood. Nonmodified synthetic mRNAs are immunogenic and activate cellular defense mechanisms, which can lead to cell death and inhibit mRNA translation upon repetitive transfection. Hence, to overcome RNA…

Homeobox protein NANOGCellular Reprogramming TechniquesInduced Pluripotent Stem CellsVaccinia virusFibroblastsBiologyTransfectionLIN28Molecular biologyCell biologyKruppel-Like Factor 4MicroRNAsSOX2KLF4GeneticsHumansMolecular MedicineCellular Reprogramming TechniquesRNA MessengerProgenitor cellInduced pluripotent stem cellMolecular BiologyReprogrammingEndothelial Progenitor CellsImmune EvasionHuman Gene Therapy
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A Trans-amplifying RNA Vaccine Strategy for Induction of Potent Protective Immunity

2019

Here, we present a potent RNA vaccine approach based on a novel bipartite vector system using trans-amplifying RNA (taRNA). The vector cassette encoding the vaccine antigen originates from an alphaviral self-amplifying RNA (saRNA), from which the replicase was deleted to form a transreplicon. Replicase activity is provided in trans by a second molecule, either by a standard saRNA or an optimized non-replicating mRNA (nrRNA). The latter delivered 10- to 100-fold higher transreplicon expression than the former. Moreover, expression driven by the nrRNA-encoded replicase in the taRNA system was as efficient as in a conventional monopartite saRNA system. We show that the superiority of nrRNA- ov…

Translational efficiencyGenetic VectorsRNA-dependent RNA polymeraseHemagglutinin (influenza)Hemagglutinin Glycoproteins Influenza VirusBiologyAntibodies ViralMadin Darby Canine Kidney CellsMice03 medical and health sciencesDogsImmunogenicity VaccineInfluenza A Virus H1N1 Subtype0302 clinical medicineOrthomyxoviridae InfectionsCricetinaeInfluenza HumanDrug DiscoveryGeneticsAnimalsHumansViral Replicase Complex ProteinsRepliconMolecular BiologyGene030304 developmental biologyPharmacologyMice Inbred BALB C0303 health sciencesMessenger RNAVaccinationRNATranslation (biology)Antibodies NeutralizingSemliki forest virusVirologyHEK293 CellsInfluenza Vaccines030220 oncology & carcinogenesisbiology.proteinRNA ViralMolecular MedicineFemaleOriginal ArticleMolecular Therapy
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Improvement of In Vivo Expression of Genes Delivered by Self-Amplifying RNA Using Vaccinia Virus Immune Evasion Proteins.

2017

Among nucleic acid–based delivery platforms, self-amplifying RNA (saRNA) vectors are of increasing interest for applications such as transient expression of recombinant proteins and vaccination. saRNA is safe and, due to its capability to amplify intracellularly, high protein levels can be produced from even minute amounts of transfected templates. However, it is an obstacle to full exploitation of this platform that saRNA induces a strong innate host immune response. In transfected cells, pattern recognition receptors sense double-stranded RNA intermediates and via activation of protein kinase R (PKR) and interferon signaling initiate host defense measures including a translational shutdow…

0301 basic medicineGenetic VectorsGene Expressionvaccinia virus E3Vaccinia virusBiologyCell Line03 medical and health sciencesMiceViral ProteinseIF-2 Kinase0302 clinical medicineImmune systemInterferonSense (molecular biology)GeneticsmedicineAnimalsHumansalphavirusMolecular BiologyResearch ArticlesImmune EvasionMessenger RNAMice Inbred BALB Cself-amplifying RNAPattern recognition receptorGene Transfer TechniquesRNAProtein kinase RVirology030104 developmental biologyvaccinia virus K3030220 oncology & carcinogenesisMolecular MedicineRNAFemalesaRNAmedicine.drugrepliconvaccinia virus B18Human gene therapy
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T-cell receptor transfer into human T cells with ecotropic retroviral vectors

2014

Adoptive T-cell transfer for cancer immunotherapy requires genetic modification of T cells with recombinant T-cell receptors (TCRs). Amphotropic retroviral vectors (RVs) used for TCR transduction for this purpose are considered safe in principle. Despite this, TCR-coding and packaging vectors could theoretically recombine to produce replication competent vectors (RCVs), and transduced T-cell preparations must be proven free of RCV. To eliminate the need for RCV testing, we transduced human T cells with ecotropic RVs so potential RCV would be non-infectious for human cells. We show that transfection of synthetic messenger RNA encoding murine cationic amino-acid transporter 1 (mCAT-1), the re…

CD4-Positive T-LymphocytesAdoptive cell transfermedicine.medical_treatmentGenetic enhancementGenetic VectorsReceptors Antigen T-CellCD8-Positive T-LymphocytesBiologyImmunotherapy AdoptiveJurkat cellsVesicular stomatitis Indiana virusCell LineJurkat CellsMiceTransduction (genetics)Viral Envelope ProteinsCancer immunotherapyTransduction GeneticGeneticsmedicineAnimalsHumansRNA MessengerMolecular BiologyCationic Amino Acid Transporter 1Membrane GlycoproteinsHEK 293 cellsT-cell receptorTransfectionAdoptive TransferVirologyElectroporationHEK293 CellsRetroviridaeLeukemia Virus Gibbon ApeMolecular MedicinePlasmidsGene Therapy
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