Difference between revisions of "Part:BBa K2206006"

Revision as of 08:47, 3 October 2017

Toehold switch for hsa-miR-15b-5p with GFPmut3b and inducible promoter

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.

This part codes for a toehold switch that contains a region that is complementary to the micro-RNA hsa-mir-15b-5p (the trigger RNA). The toehold switch is activated by hsa-mir-15b-5p and regulates production of GFPmut3b. Fluorescence intensity of GFP is proportional to the number of toehold switches activated, thus indicating the levels of hsa-mir-15b-5p present. This part can therefore be used to quantify the levels of hsa-mir-15b-5p.

We believe that the transcripts produced from this part are toxic when expressed at reasonably high concentrations in E.coli (e.g. the concentrations produced from the BBa_J23111 constitutive promoter). An iducible promoter is therefore used to allow for amplification of the constructs in E.coli.

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

Sequence and Features