Composite

Part:BBa_K3328018:Design

Designed by: Qi Wu   Group: iGEM20_OUC-China   (2020-10-04)


triggers of IMPLY gate (IMPLY1)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We combined the 3WJ switch and toehold switch to realize the IMPLY Boolean calculation. When no trigger is expressed, this logic gate just likes a 3WJ switch. When trigger A expressed, the trigger will bind to the switch RNA. The binding allows for a branch migration process, exposing AUG and RBS for translation initiation. When trigger B expressed, the trigger will bind to 3WJ switch RNA. The resulting trigger–switch complex has a stable 3WJ structure that effectively sequesters the RBS and the start codon within the loop and stem of the switch RNA, respectively, and strongly represses translation. When both trigger A and B are expressed, the binding of trigger RNA to the toehold sequence allows the RNA polymerase to bind to the former RBS and break open 3WJ stable hairpin.

799px-T--OUC-China--design_lunbo_imply.jpg

Source

synthesize from company


References

[1] Kim, J., Zhou, Y., Carlson, P. D., Teichmann, M., Chaudhary, S., Simmel, F. C., … Green, A. A. (2019). De novo-designed translation-repressing riboregulators for multi-input cellular logic. Nature Chemical Biology. doi:10.1038/s41589-019-0388-1

[2] Green, A. A., Silver, P. A., Collins, J. J., and Yin, P. (2014) toehold switches: de-novo- designed regulators of gene expression. Cell 159, 925– 939, DOI: 10.1016/j.cell.2014.10.002