Regulatory

Part:BBa_K2206010

Designed by: Sammy Lovat   Group: iGEM17_CLSB-UK   (2017-10-18)
Revision as of 19:52, 18 October 2017 by Registry (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


Toehold switch for hsa-miR-27b-3p (second series)

Toehold switches are synthetic riboregulators that regulate gene expression post-transcriptionally. Gene expression can be activated in the presence of a cognate single stranded RNA molecule that contains an arbitrary sequence (the trigger RNA). The trigger RNA binds to the switch through base pairing, causing a conformational change that results in translation of the downstream protein coding region.

The trigger RNA sequence can be separated into different RNA molecules. Hybridisation of these different RNA molecules results in a complete trigger RNA sequence, that is capable of activating the toehold switch.

Bba_X and hsa-miR-15b-5p contain roughly half of the trigger sequence each. Colocalization of the split trigger RNA sequences results in a functional trigger RNA that is capable of activating the toehold switch. This part codes for a toehold switch that contains a region that is complementary to the complete trigger RNA sequence formed of Bba_X and hsa-miR-15b-5p Therefore, this part can be used to regulate the expression of any protein in response to presence of both Bba_X and hsa-miR-15b-5p.

We found that the transcripts produced by our other toehold switch parts were toxic to E.coli. We therefore recommend using this part with a non-leaky promoter (for reference, we found K808000 to be too leaky).

N.B. This part contains a strong RBS sequence.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 65


[edit]
Categories
Parameters
None