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Engineering mRNA Untranslated Region Structure to Regulate Translation in Mammalian Cells

Zhang, Heyan LU (2026) KBTM01 20261
Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
Abstract
Messenger RNA (mRNA) plays a central role in gene expression by transferring genetic information from DNA to the translational machinery for protein synthesis. Recent advances in RNA therapeutics and synthetic biology have highlighted the importance of untranslated regions (UTRs) and RNA secondary structures in regulating mRNA stability and translational efficiency. In the present study, engineered mRNA constructs containing designed untranslated regions with distinct predicted secondary structures were investigated in a mammalian in vitro transcribed mRNA system.
Synthetic mRNA constructs encoding green fluorescent protein (GFP) were computationally designed. DNA templates containing engineered UTRs and a T7 promoter were generated by... (More)
Messenger RNA (mRNA) plays a central role in gene expression by transferring genetic information from DNA to the translational machinery for protein synthesis. Recent advances in RNA therapeutics and synthetic biology have highlighted the importance of untranslated regions (UTRs) and RNA secondary structures in regulating mRNA stability and translational efficiency. In the present study, engineered mRNA constructs containing designed untranslated regions with distinct predicted secondary structures were investigated in a mammalian in vitro transcribed mRNA system.
Synthetic mRNA constructs encoding green fluorescent protein (GFP) were computationally designed. DNA templates containing engineered UTRs and a T7 promoter were generated by PCR amplification and plasmid construction, followed by in vitro transcription (IVT) to produce capped and polyadenylated mRNAs. The resulting IVT-generated mRNAs were directly transfected into HEK293T cells, and GFP expression was evaluated using confocal fluorescence microscopy.
Differences in GFP fluorescence intensity were observed among target RNAs, suggesting that RNA secondary structure within untranslated regions can influence translational efficiency in mammalian cells. These findings support previous studies demonstrating that RNA accessibility and structural stability play important roles in translation initiation and protein production.
In addition to experimental evaluation, the study discusses the limitations of computational RNA structure prediction and the importance of future approaches such as RT-qPCR and programmable RNA regulatory systems for more comprehensive characterization of translational control mechanisms. Overall, this work provides a framework for investigating structure–function relationships in synthetic mRNA systems and may contribute to future optimization of RNA-based imaging and synthetic biology applications. (Less)
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author
Zhang, Heyan LU
supervisor
organization
course
KBTM01 20261
year
type
H2 - Master's Degree (Two Years)
subject
keywords
mRNA, untranslated regions, UTRs, RNA secondary structure, in vitro transcription, translation efficiency, biotechnology
language
English
id
9235440
date added to LUP
2026-08-11 09:11:36
date last changed
2026-08-11 09:11:36
@misc{9235440,
  abstract     = {{Messenger RNA (mRNA) plays a central role in gene expression by transferring genetic information from DNA to the translational machinery for protein synthesis. Recent advances in RNA therapeutics and synthetic biology have highlighted the importance of untranslated regions (UTRs) and RNA secondary structures in regulating mRNA stability and translational efficiency. In the present study, engineered mRNA constructs containing designed untranslated regions with distinct predicted secondary structures were investigated in a mammalian in vitro transcribed mRNA system.
Synthetic mRNA constructs encoding green fluorescent protein (GFP) were computationally designed. DNA templates containing engineered UTRs and a T7 promoter were generated by PCR amplification and plasmid construction, followed by in vitro transcription (IVT) to produce capped and polyadenylated mRNAs. The resulting IVT-generated mRNAs were directly transfected into HEK293T cells, and GFP expression was evaluated using confocal fluorescence microscopy.
Differences in GFP fluorescence intensity were observed among target RNAs, suggesting that RNA secondary structure within untranslated regions can influence translational efficiency in mammalian cells. These findings support previous studies demonstrating that RNA accessibility and structural stability play important roles in translation initiation and protein production.
In addition to experimental evaluation, the study discusses the limitations of computational RNA structure prediction and the importance of future approaches such as RT-qPCR and programmable RNA regulatory systems for more comprehensive characterization of translational control mechanisms. Overall, this work provides a framework for investigating structure–function relationships in synthetic mRNA systems and may contribute to future optimization of RNA-based imaging and synthetic biology applications.}},
  author       = {{Zhang, Heyan}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Engineering mRNA Untranslated Region Structure to Regulate Translation in Mammalian Cells}},
  year         = {{2026}},
}