IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v16y2025i1d10.1038_s41467-025-62588-y.html
   My bibliography  Save this article

Simultaneous in vitro expression of minimal 21 transfer RNAs by tRNA array method

Author

Listed:
  • Ryota Miyachi

    (The University of Tokyo)

  • Keiko Masuda

    (Chuo-ku)

  • Yoshihiro Shimizu

    (Chuo-ku)

  • Norikazu Ichihashi

    (The University of Tokyo
    The University of Tokyo
    The University of Tokyo)

Abstract

Transfer RNA (tRNA) plays a central role in translation. The simultaneous in vitro synthesis of minimal yet sufficient tRNA species (at least 21) poses a challenge for constructing a self-reproducible artificial cell. A key obstacle is the processing of the 5’ and 3’ ends, which requires a multi-step reaction in the natural cells. In this study, we develop a simplified processing method that allows simultaneous expression of all 21 tRNAs in a reconstituted transcription/translation system (PURE system). We test two methods for 5’-end processing (the leader and 5’-G variants methods) and two methods for 3’-end processing (the direct tRNA linkage and HDVR attachment methods). Finally, by combining the direct tRNA linkage and HDVR attachment methods (newly termed the tRNA array method), we succeed in simultaneously expressing all 21 tRNAs from a single polycistronic DNA template in the PURE system. The tRNA mixture produced by the tRNA array method supports a similar level of translation to the individually synthesized tRNA mixture for luciferase and GFP. This study represents a step toward the realization of self-reproducible artificial cells and also provides an easy method for preparing all tRNAs useful for genetic code engineering.

Suggested Citation

  • Ryota Miyachi & Keiko Masuda & Yoshihiro Shimizu & Norikazu Ichihashi, 2025. "Simultaneous in vitro expression of minimal 21 transfer RNAs by tRNA array method," Nature Communications, Nature, vol. 16(1), pages 1-17, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-62588-y
    DOI: 10.1038/s41467-025-62588-y
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-025-62588-y
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-025-62588-y?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-62588-y. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.