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mRNA structure regulates protein expression through changes in functional half-life.

David M Mauger1, B Joseph Cabral1, Vladimir Presnyak1

  • 1Platform Research, Moderna, Inc., Cambridge, MA 02139.

Proceedings of the National Academy of Sciences of the United States of America
|November 13, 2019
PubMed
Summary
This summary is machine-generated.

Messenger RNA (mRNA) structure significantly impacts protein expression. Stabilizing mRNA secondary structure with modified nucleotides enhances protein production, revealing its crucial role in mRNA stability and translation.

Keywords:
RNA structureSHAPEmRNA therapueticsmodified nucleotidestranslation

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Messenger RNAs (mRNAs) contain regulatory information in both their sequence and higher-order structure.
  • The distinct roles of codon usage and mRNA secondary structure in protein expression are challenging to isolate due to their correlation in natural sequences.

Purpose of the Study:

  • To investigate the independent contribution of messenger RNA (mRNA) secondary structure to protein expression regulation.
  • To differentiate the effects of mRNA structure from codon usage on protein synthesis.

Main Methods:

  • Utilized global inclusion of modified nucleotides to stabilize mRNA secondary structure.
  • Employed rational sequence design of exogenous mRNA constructs to manipulate coding sequence (CDS) structure independently of codon usage.
  • Assessed protein expression levels and functional mRNA half-life using enhanced green fluorescent protein (eGFP) mRNA variants.

Main Results:

  • Highly expressed mRNAs were found to possess highly structured coding sequences (CDS).
  • Modified nucleotides that enhance mRNA secondary structure promoted high protein expression across diverse sequences.
  • Altering CDS structure, independent of codon usage, directly impacted protein expression by modulating functional mRNA half-life.

Conclusions:

  • mRNA secondary structure plays a critical, underappreciated role in regulating protein expression.
  • Stabilizing mRNA secondary structure is a viable strategy for enhancing protein production.
  • Functional mRNA half-life, defined by active translation, is modulated by mRNA CDS structure.