Difference between revisions of "Part:BBa K2206003"
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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.
 
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.
  
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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.  
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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-27b-3p 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-27b-3p present.  
  
 
This part can therefore be used to quantify the levels of hsa-mir-15b-5p.
 
This part can therefore be used to quantify the levels of hsa-mir-15b-5p.

Revision as of 22:45, 2 October 2017


Toehold switch for hsa-miR-27b-3p with GFPmut3b

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-27b-3p 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-27b-3p 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).

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

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 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 758