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A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
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Non-viral 2A-like sequences for protein coexpression.

Wei Wen Su1, Bei Zhang1, Zhenlin Han1

  • 1Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.

Journal of Biotechnology
|August 22, 2022
PubMed
Summary

Researchers explored non-viral 2A-like peptides from sea urchin and sea slug for polyprotein coexpression. These novel sequences function like viral 2A peptides, enabling protein production without animal-virus components.

Keywords:
Medical biotechnologyPlant biotechnologyProtein expressionRecombinant antibodySynthetic biology

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Area of Science:

  • Biotechnology and synthetic biology
  • Molecular and cellular biology

Background:

  • Simultaneous coexpression of multiple proteins is crucial for biotechnology and synthetic biology.
  • The foot-and-mouth disease virus (FMDV) 2A peptide is widely used for polyprotein coexpression via translational ribosome skipping.
  • Consumer concerns regarding transgenic products with animal-virus sequences necessitate alternative coexpression strategies.

Purpose of the Study:

  • To investigate novel non-viral 2A-like peptides from marine invertebrates for polyprotein coexpression.
  • To evaluate the efficacy of these non-viral peptides as alternatives to viral 2A peptides.
  • To demonstrate their utility in producing functional proteins, such as recombinant antibodies.

Main Methods:

  • Identification and characterization of 2A-like sequences from purple sea urchin (Strongylocentrotus purpuratus) and California sea slug (Aplysia californica).
  • Testing the polyprotein processing activity of these non-viral sequences in both plant and mammalian cell systems.
  • Assessing the functionality of proteins produced using these novel coexpression systems, including a recombinant antibody.

Main Results:

  • The non-viral 2A-like peptides from sea urchin and sea slug effectively mediated polyprotein processing, similar to the viral FMDV 2A peptide.
  • Successful coexpression and processing were achieved in both plant and mammalian cells.
  • A functional recombinant antibody was successfully produced using these novel non-viral coexpression systems.

Conclusions:

  • Non-viral 2A-like peptides from marine invertebrates offer a viable and safe alternative to viral 2A peptides for polyprotein coexpression.
  • These novel sequences can be employed in diverse cell systems (plant and mammalian) for producing multiple proteins simultaneously.
  • This advancement provides a valuable tool for engineering multigenic traits and producing protein complexes for biomedical applications, addressing consumer concerns about animal-virus components.