Mechanistic modeling of in vitro transcription incorporating effects of magnesium pyrophosphate crystallization

Stover, Nathan Merica; Ganko, Krystian; Braatz, Richard D.

doi:10.1002/bit.28699

Title: Mechanistic modeling of in vitro transcription incorporating effects of magnesium pyrophosphate crystallization

Journal Article · 02 May 2024 · Biotechnology and Bioengineering

DOI: https://doi.org/10.1002/bit.28699 · OSTI ID:2345234

Stover, Nathan Merica ^[1]; Ganko, Krystian ^[1];

^[1]

Department of Chemical Engineering Massachusetts Institute of Technology Cambridge MA USA

Abstract The in vitro transcription (IVT) reaction used in the production of messenger RNA vaccines and therapies remains poorly quantitatively understood. Mechanistic modeling of IVT could inform reaction design, scale‐up, and control. In this work, we develop a mechanistic model of IVT to include nucleation and growth of magnesium pyrophosphate crystals and subsequent agglomeration of crystals and DNA. To help generalize this model to different constructs, a novel quantitative description is included for the rate of transcription as a function of target sequence length, DNA concentration, and T7 RNA polymerase concentration. The model explains previously unexplained trends in IVT data and quantitatively predicts the effect of adding the pyrophosphatase enzyme to the reaction system. The model is validated on additional literature data showing an ability to predict transcription rates as a function of RNA sequence length.

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Sponsoring Organization:: USDOE

OSTI ID:: 2345234

Alternate ID(s):: OSTI ID: 2429466

Journal Information:: Biotechnology and Bioengineering, Journal Name: Biotechnology and Bioengineering Vol. 121 Journal Issue: 9; ISSN 0006-3592

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: United States

Language:: English

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