Methods of Higher Fidelity RNA Synthesis

Abstract: the present disclosure relates, according to some embodiments, to methods and compositions for improving fidelity of nucleotide incorporation during rna synthesis. immunogenicity of in vitro transcribed synthetic mrnas may be reduced and/or therapeutic efficacy may be increased by rna sequence optimization and/or incorporation of modified uridine analogs (e.g. pseudouridine (�) and n-methyl-pseudouridine (m1�)). the present disclosure relates to protocols to improve uridine analog incorporation-without affecting total rna yield-during ivt. methods for higher-fidelity incorporation of uridine analogs during ivt include guidelines when choosing ssrnaps for the generation of modified uridine-containing mrnas in vitro.

Inventor(s): Bijoyita Roy, Tien-Hao Chen, Vladimir Potapov, Jennifer Ong, Nan Dai

CPC Classification: C12P19/34 (FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer ; producing vinegar ; producing specific peptides or proteins ; producing enzymes ; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification ; measuring or testing processes involving enzymes or microorganisms ; measuring or testing processes involving nucleic acid amplification reactions ; fermentation processes to form a food composition, or ; compounds in general, see the relevant compound class, e.g. , )})

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