The invention relates to a method of quantifying capping efficiency in a sample from an in vitro transcription reaction mixture comprising a plurality of mRNA transcript, characterized by a step of contacting the mRNA transcripts with an oligonucleotide complementary to a sequence of nucleotides in the...
Maximizing the production of therapeutic proteins from mRNA medicines requires a thorough understanding of how nucleotide sequence, nucleotide modification and RNA structure interplay to affect translational efficiency and mRNA stability. In this Review, we describe the principles that underlie the physical ...
Capping EfficiencyLC-MS≥ 90% Size based purityCapillary Electrophoresis≥75% ImpurityTotal protein residueNano Orange Assay≤ 1% Plasmid DNA ResidueqPCR≤ 0.05% dsRNASlot-blot≤ 0.1% Aggregate quantificationRNA Aggregation Assay<5% SafetyEndotoxinSemiquantitative< 10EU/mg ...
Maximizing the production of therapeutic proteins from mRNA medicines requires a thorough understanding of how nucleotide sequence, nucleotide modification and RNA structure interplay to affect translational efficiency and mRNA stability. In this Review, we describe the principles that underlie the physical ...
N1-methylpseudouridine (see ‘Methods’). We prepared both short- and long-read sequencing libraries from the modified mRNA. These libraries had lower yields (~50%) than matched, native mRNA vaccines, suggesting the modified nucleosides reduced the efficiency of cDNA library preparation (Fig.S9d...
Realizing the immense clinical potential of mRNA-based drugs will require continued development of methods to safely deliver the bioactive agents with high efficiency and without triggering side effects. In this regard, lipid nanoparticles have been succ
Efficiency of splicing mRNAs提高剪接效率 2 3'加尾(多腺苷酸化) hnRNA(heterogeneous nuclear mRNA)是mRNA的前体,它们的3'端有一长串AMP残基,这被称为poly(A)尾。tRNA和rRNA则都没有。 将poly(A)尾加入RNA的过程称为多腺苷酸化(polyadenylation)。
At, we believe that the best practice approach to supporting the analytical development of mRNA therapies involves using orthogonal techniques wherever possible. Methods should be validated appropriately to the relevant phase of clinical development, regulatory submission requirements, or for supporting ongoin...
translational efficiency and uptake of mRNA. These optimizations were achieved by modification of its non-coding elements (5′ cap structure and its capping efficiency [1,2,3,4], 5′- and 3′-untranslated regions (UTRs) [5,6,7,8,9], 3′ poly(A) tail [5,10,11]) and of the codin...
enzyme capping method. The capping efficiency of ARCA is relatively low (70%), and it produces an immunogenic cap 0 structure. However, in the two-step capping method using vaccinia virus-derived capping enzyme, capping efficiency is almost 100% (Martin et al.1975). Eukaryotic mRNA has severa...