Imperial College London
Alternate

Dr Paul F. McKay

Faculty of Medicine, Department of Infectious Disease

Advanced Research Fellow
 
 
 
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Contact

 

+44 (0)20 7594 2542p.mckay

 
 
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Location

 

125 (Shattock Group)Wright Fleming WingSt Mary's Campus

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Summary

 

Publications

Citation

BibTex format

@unpublished{Samnuan:2021:10.1101/2021.01.08.425833,
author = {Samnuan, K and Blakney, A and McKay, P and Shattock, R},
doi = {10.1101/2021.01.08.425833},
publisher = {Cold Spring Harbor Laboratory},
title = {Design-of-experiments in vitro transcription yield optimization of self-amplifying RNA},
url = {http://dx.doi.org/10.1101/2021.01.08.425833},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - UNPB
AB  - Self-amplifying RNA (saRNA) vaccines are able to induce a higher antigen-specific immune response with a more cost-effective and rapid production process compared to plasmid DNA vaccines. saRNAs are synthesized through in vitro transcription (IVT) however; this process has mainly been optimized for relatively short mRNAs. Here, we optimized the IVT process for long saRNAs, approximately 9.4 kb through a design of experiment (DoE) approach to produce a maximal RNA yield and validated the optimal IVT method on various sizes of RNA. We found that magnesium has the highest impact on RNA yield with acetate ions enabling a higher yield than chloride ions. In addition, the interaction between magnesium and nucleoside triphosphates (NTPs) is highly essential for IVT. Further addition of sodium acetate (NaOAc) during IVT provided no added benefit in RNA yield. Moreover, pyrophosphatase was not essential for productive IVT. The optimal IVT method can be used to synthesize different lengths of RNA. These findings emphasize the ability to synthesize high quality and quantity of saRNA through IVT and that the optimal amount of each component is essential for their interactions to produce a high RNA yield.
AU  - Samnuan,K
AU  - Blakney,A
AU  - McKay,P
AU  - Shattock,R
DO  - 10.1101/2021.01.08.425833
PB  - Cold Spring Harbor Laboratory
PY  - 2021///
TI  - Design-of-experiments in vitro transcription yield optimization of self-amplifying RNA
UR  - http://dx.doi.org/10.1101/2021.01.08.425833
UR  - https://www.biorxiv.org/content/10.1101/2021.01.08.425833v1
UR  - http://hdl.handle.net/10044/1/94974
ER  -
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