Coding

Part:BBa_K3727006

Designed by: Danai Eleni Vergini   Group: iGEM21_Thessaloniki   (2021-09-07)


Toehold_Switch-30e-5p (3)

This part is a toehold switch, an RNA molecule that includes two crucial areas. The first one is complementary to the sequence of the miRNA molecule hsa-miR-30e-5p. The second one is complementary to a different part of the molecule, so that alongside sequences of our own design , form a hairpin-structure. A Ribosome Binding Site (RBS) is embedded in this structure, so that the binding of the ribosome and as a result the translation of the downstream genes is inhibited. This molecule can be linked to a reporter gene (such as GFP, BFP, mCherry etc.) to regulate its expression. In the presence of hsa-miR-30e-5p which is complementary to the first part of the Toehold_Switch-30e-5p (3) the hairpin unfolds and the ribosome is able to bind to the RBS region and the expression of the reporter is initiated [1],[2].

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] A. A. Green, P. A. Silver, J. J. Collins and P. Yin, “Toehold switches: De-novo-designed regulators of gene expression”, Cell, vol. 159, no. 4, pp 925–939, Oct. 2014, https://doi.org/10.1016/j.cell.2014.10.002.

[2] K. Pardee, A. A. Green, M. K. Takahashi, D. Braff, G. Lambert, J. W. Lee, T. Ferrante, D. Ma, N. Donghia, M. Fan, N. M. Daringer, I. Bosch, D. M. Dudley, D. H. O’Connor, L. Gehrke and J. J. Collins, “Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components”, Cell, vol. 165, no. 5, pp 1255–1266, May 2016, https://doi.org/10.1016/j.cell.2016.04.059.

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