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Revision as of 12:37, 1 October 2023


Twister P1 ribozyme, from 10.1080/15476286.2022.2123640 contributed by Fudan iGEM 2023


Introduction

The twister ribozyme is an RNA molecule that is capable of self-cleavage at specific sites.The RNA exhibits nucleolytic ribozyme activity both in vitro and in vivo, with cleavage rates comparable to those of other nucleolytic ribozymes[1]. This grants it the ability to regulate polycistronic co-expression system[2]

Usage and Biology

The Twister ribozyme is a crucial component of our ribozyme-assisted polycistronic co-expression system：pRAP.

We incorporate the twister ribozyme between the coding CDSs in a polycistron. This ribozyme cleaves the polycistronic mRNA transcript, converting them into individual mono-cistrons. This process eliminates interactions between the cistrons, ensuring each mono-cistron has a comparable translation initiation rate. Subsequently, in conjunction with the RBS designed by the pRAPer software, we can modulate the translation initiation rate of individual mono-cistrons, enabling precise adjustment of the pRAP co-expression system.

Characterization

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. ↑ Eiler, D., Wang, J., & Steitz, T. A. (2014). Structural basis for the fast self-cleavage reaction catalyzed by the twister ribozyme. Proceedings of the National Academy of Sciences, 111(36), 13028–13033. https://doi.org/10.1073/pnas.1414571111
  2. ↑ Liu, Y., Wu, Z., Wu, D., Gao, N., & Lin, J. (2022). Reconstitution of Multi-Protein Complexes through Ribozyme-Assisted Polycistronic Co-Expression. ACS Synthetic Biology, 12(1), 136–143. https://doi.org/10.1021/acssynbio.2c00416