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Related Concept Videos

Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

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Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
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Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

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The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
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What is Gene Expression?01:36

What is Gene Expression?

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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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Regulated mRNA Transport02:22

Regulated mRNA Transport

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In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing...
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Nuclear Export of mRNA02:31

Nuclear Export of mRNA

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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

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The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
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Reliably Engineering and Controlling Stable Optogenetic Gene Circuits in Mammalian Cells
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Design of a self-regulating mRNA gene circuit.

Eric C Dykeman1

  • 1Department of Mathematics, University of York, York, YO10 5DD, UK. eric.dykeman@york.ac.uk.

Scientific Reports
|August 21, 2024
PubMed
Summary

Researchers designed self-regulating messenger RNA (mRNA) molecules that control protein production without affecting transcription. This breakthrough enables precise protein level control for synthetic biology and vaccine applications.

Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Virology

Background:

  • In vivo protein expression is typically regulated by transcriptional feedback mechanisms.
  • Positive-sense single-stranded RNA viruses often control protein expression through mRNA secondary structures like internal ribosomal entry sites.
  • This viral self-regulation suggests potential for designing artificial self-regulating mRNAs.

Purpose of the Study:

  • To demonstrate the feasibility of designing self-regulating messenger RNA (mRNA) molecules.
  • To create a proof-of-concept polycistronic mRNA system for controlled protein expression.
  • To explore applications in synthetic biology and vaccine development requiring precise protein level control.

Main Methods:

  • Designed a polycistronic mRNA based on bacteriophage MS2.

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  • Engineered an upstream gene to repress the synthesis of a downstream gene.
  • Utilized a computational tool to simulate ribosome kinetics and co-translational mRNA folding.

    • Main Results:

    • Identified specific mRNA mutations that enhance translation efficiency.
    • Demonstrated that mutations can improve the repression of downstream gene expression.
    • Validated the concept of self-regulating mRNA for controlling protein synthesis.

    Conclusions:

    • Self-regulating mRNA gene circuits can be designed for precise control of cellular protein levels.
    • This approach offers a novel strategy for developing mRNA-based therapeutics and vaccines.
    • The findings open possibilities for bespoke mRNA engineering in synthetic biology.