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Engineered ribosomes with tethered subunits for expanding biological function

Author

Listed:
  • Erik D. Carlson

    (Northwestern University
    Northwestern University
    Northwestern University
    Stanford University)

  • Anne E. d’Aquino

    (Northwestern University
    Northwestern University
    Northwestern University
    Northwestern University)

  • Do Soon Kim

    (Northwestern University
    Northwestern University
    Northwestern University)

  • Emily M. Fulk

    (Northwestern University
    Northwestern University
    Northwestern University)

  • Kim Hoang

    (Northwestern University
    Johnson and Wales University)

  • Teresa Szal

    (University of Illinois at Chicago)

  • Alexander S. Mankin

    (University of Illinois at Chicago)

  • Michael C. Jewett

    (Northwestern University
    Northwestern University
    Northwestern University
    Northwestern University)

Abstract

Ribo-T is a ribosome with covalently tethered subunits where core 16S and 23S ribosomal RNAs form a single chimeric molecule. Ribo-T makes possible a functionally orthogonal ribosome–mRNA system in cells. Unfortunately, use of Ribo-T has been limited because of low activity of its original version. Here, to overcome this limitation, we use an evolutionary approach to select new tether designs that are capable of supporting faster cell growth and increased protein expression. Further, we evolve new orthogonal Ribo-T/mRNA pairs that function in parallel with, but independent of, natural ribosomes and mRNAs, increasing the efficiency of orthogonal protein expression. The Ribo-T with optimized designs is able to synthesize a diverse set of proteins, and can also incorporate multiple non-canonical amino acids into synthesized polypeptides. The enhanced Ribo-T designs should be useful for exploring poorly understood functions of the ribosome and engineering ribosomes with altered catalytic properties.

Suggested Citation

  • Erik D. Carlson & Anne E. d’Aquino & Do Soon Kim & Emily M. Fulk & Kim Hoang & Teresa Szal & Alexander S. Mankin & Michael C. Jewett, 2019. "Engineered ribosomes with tethered subunits for expanding biological function," Nature Communications, Nature, vol. 10(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-11427-y
    DOI: 10.1038/s41467-019-11427-y
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    Cited by:

    1. Wenwen Yu & Ke Jin & Dandan Wang & Nankai Wang & Yangyang Li & Yanfeng Liu & Jianghua Li & Guocheng Du & Xueqin Lv & Jian Chen & Rodrigo Ledesma-Amaro & Long Liu, 2024. "De novo engineering of programmable and multi-functional biomolecular condensates for controlled biosynthesis," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    2. Antje Krüger & Andrew M. Watkins & Roger Wellington-Oguri & Jonathan Romano & Camila Kofman & Alysse DeFoe & Yejun Kim & Jeff Anderson-Lee & Eli Fisker & Jill Townley & Anne E. d’Aquino & Rhiju Das & , 2023. "Community science designed ribosomes with beneficial phenotypes," Nature Communications, Nature, vol. 14(1), pages 1-11, December.
    3. Roberto Di Blasi & Mara Pisani & Fabiana Tedeschi & Masue M. Marbiah & Karen Polizzi & Simone Furini & Velia Siciliano & Francesca Ceroni, 2023. "Resource-aware construct design in mammalian cells," Nature Communications, Nature, vol. 14(1), pages 1-10, December.

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