About this Research Topic
The COVID-19 pandemic triggered an unprecedented rate of development of mRNA vaccines, and recently new RNA-based therapeutic modalities, such as self-amplifying RNA and circular RNA. Production of RNA therapeutics typically involves 10-15 steps including plasmid production, plasmid linearization, in vitro transcription (IVT) reaction, purification, and encapsulation. Although the manufacturing processes vary significantly, the common unit of operation is the IVT reaction, which produces RNA from a DNA template. Despite its central role in RNA production, it has received relatively little academic attention. To advance our understanding of factors influencing both the quality and yield of RNA, new approaches to understanding IVT kinetics are needed, as well as methodologies to either improve or obviate the need for 5’ capping and 3’ tailing. Novel modalities pose significant purification challenges, which should also be considered.
The goal of the current Research Topic is to cover promising, recent, and novel research trends in the molecular understanding of RNA synthesis for therapeutic applications. Areas to be covered in this Research Topic may include, but are not limited to:
Towards a molecular understanding of IVT reaction:
o kinetic studies of polymerase activity, including inhibition mechanisms
o new analytical tools to study IVT (spectroscopic, HPLC, MS)
o novel polymerase designs (e.g. thermophilic, halophilic, etc)
o DoE approaches to IVT optimization (increasing yield, decreasing immunogenicity)
o Modelling approaches for continuous RNA production
Optimization and understanding of 5' capping:
o novel co-transcriptional cap analogues
o advances in enzymatic capping
o molecular biology approaches to produce capped RNA (e.g. mRNA production in eukaryotic cells)
o improving analytics of 5' capping: molecular biology methods to determine 5' capping, high-throughput methods, MS-based methods
o uncapped RNA therapeutics
Novel therapeutic modalities produced by IVT
o Utilizing molecular understanding of IVT to produce novel RNA modalities: circular RNA, self-amplifying RNA, self-replicating RNA
o Addressing purification challenges for new RNA-based therapeutic modalities, especially non-affinity purification methodologies
Dr Sekirnik and Dr Kuhn are employees of Sartorius BIA Separations and BioNTech, respectively. All other editors declare no conflict of interest
The goal of the current Research Topic is to cover promising, recent, and novel research trends in the molecular understanding of RNA synthesis for therapeutic applications. Areas to be covered in this Research Topic may include, but are not limited to:
Towards a molecular understanding of IVT reaction:
o kinetic studies of polymerase activity, including inhibition mechanisms
o new analytical tools to study IVT (spectroscopic, HPLC, MS)
o novel polymerase designs (e.g. thermophilic, halophilic, etc)
o DoE approaches to IVT optimization (increasing yield, decreasing immunogenicity)
o Modelling approaches for continuous RNA production
Optimization and understanding of 5' capping:
o novel co-transcriptional cap analogues
o advances in enzymatic capping
o molecular biology approaches to produce capped RNA (e.g. mRNA production in eukaryotic cells)
o improving analytics of 5' capping: molecular biology methods to determine 5' capping, high-throughput methods, MS-based methods
o uncapped RNA therapeutics
Novel therapeutic modalities produced by IVT
o Utilizing molecular understanding of IVT to produce novel RNA modalities: circular RNA, self-amplifying RNA, self-replicating RNA
o Addressing purification challenges for new RNA-based therapeutic modalities, especially non-affinity purification methodologies
Dr Sekirnik and Dr Kuhn are employees of Sartorius BIA Separations and BioNTech, respectively. All other editors declare no conflict of interest
Keywords: IVT, In vitro, Transcription, RNA, mRNA, Tools, Methods
Important Note: All contributions to this Research Topic must be within the scope of the section and journal to which they are submitted, as defined in their mission statements. Frontiers reserves the right to guide an out-of-scope manuscript to a more suitable section or journal at any stage of peer review.
