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There is widespread interest in chemically modified mRNA (modRNA) synthesized by in vitro transcription (IVT) reactions, wherein one or more of the natural (aka wild-type) A, G, C, and U nucleotide 5’-triphosphates is replaced by base-modified analogs.
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CRISPR-based genome editing tools show tremendous potential as therapeutic agents for the treatment of genetic diseases and viral infections, as discussed in previous Zone blogs and the June 2021 TriLink Research Spotlight titled, A Novel Base-Editing Strategy Promises to Treat Sickle Cell Disease . Nevertheless, safe and efficient delivery of CRISPR tools to target tissues continues to attract growing attention (see Chart). A wide variety of technical approaches are being exp...
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This post is Part 2 of a series of Zone blogs featuring DNA plasmids used to produce in vitro transcribed (IVT) mRNA. Part 1 , posted on June 8, 2021, provided historical perspectives on the discovery of plasmids, as well as descriptions of how these circular double-stranded DNAs enabled recombinant DNA technology and now IVT mRNA production. The two growth phases of plasmid applications are reflected in this chart of publications in PubMed indexed to gene therapy and genetic vacci...
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Introduction Synthetic mRNA produced by in vitro transcription (IVT) is the active pharmaceutical ingredient of approved vaccines and of many drugs under development. IVT mRNA typically contains several hundred bases of non-coding untranslated regions (UTRs) that are involved in the stabilization and translation of the mRNA. Remarkably, new research has discovered that these UTRs can be virtually eliminated without compromising translation of the encoded protein of interest. ...
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TriLink recently introduced plasmid manufacturing services as advantageous entry points for the in vitro transcription (IVT) template production of your mRNA, and this prompted the Zone to provide a series of blogs exploring plasmid biology and manufacturing. This initial (Part 1) blog offers some introductory comments on the discovery of plasmids that paved the way for recombinant DNA and early IVT mRNA technologies. Part 2, which will be posted in July of 2021, transitions to d...
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This blog provides some perspective on published strategies for this type of sequence optimization, which is alternatively referred to sequence engineering. This topic has been trending upwards in recent years, as indicated by the linear forecast dashed line shown in this chart reports found by word searching the NIH PubMed database for 2013 – 2020.
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Posted in the Zone on May 25, 2021 , part 1 of this series on optimizing the performance of in vitro translated (IVT) mRNA using N1-methylpseudouridine (N1mΨ) provided a historical introduction to the development of this chemically modified base. Part 1 focused on a 2015 study and an initial 2018 in vivo optimization study of N1mΨ-mRNA. This Part 2 blog features a subsequent 2019 publication on in vitro optimization of N1mΨ-mRNA for future heart gene therapy. Befor...
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