Mechanistic modeling of in vitro transcription incorporating effects of magnesium pyrophosphate crystallization
Name
Biotech Bioengineering - 2024 - Stover - Mechanistic modeling of in vitro transcription incorporating effects of.pdf
Description
Published version
Size
1.43 MB
Format
Adobe PDF
Checksum (MD5)
81378d3f9d94b249ff137c30a334deab
Author(s)
Stover, Nathan Merica
Ganko, Krystian
Braatz, Richard D
Date Issued
September 2024
Journal
Biotechnology and Bioengineering
Publisher
Wiley
Citation
Stover, Nathan Merica, Ganko, Krystian and Braatz, Richard D. 2024. "Mechanistic modeling of in vitro transcription incorporating effects of magnesium pyrophosphate crystallization." Biotechnology and Bioengineering, 121 (9).
Version
Final published version
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.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs
Persistent DSpace Link
DOI of Published Version
10.1002/bit.28699
