Difference between revisions of "Part:BBa K2206004"
Line 5: Line 5:
 
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.
  
−
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 Luciferase. The luminescence intensity of Luciferase is proportional to the number of toehold switches activated (as the more switches activated, the greater the amount of luciferase is produced), thus indicating the levels of hsa-mir-15b-5p present (as the more micro-RNA there is, the greater the number of switches activated). This part can therefore be used to quantify the levels of hsa-mir-15b-5p.   
+
This part codes for a toehold switch that contains a region that is complementary to the microRNA hsa-miR-15b-5p (the trigger RNA). The toehold switch is activated by hsa-miR-15b-5p and regulates production of Luciferase. The luminescence intensity of Luciferase is proportional to the number of toehold switches activated (as the more switches activated, the greater the amount of luciferase is produced), thus indicating the levels of hsa-miR-15b-5p present (as the more micro-RNA there is, the greater the number of switches activated). 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).
+
The transcripts produced by this part are toxic to E. coli. We recommend the use of a non-leaky promoter (for reference, we found BBa_K808000 was too leaky).
  
−
N.B. This part contains a strong RBS sequence.
+
This part contains a strong RBS sequence.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 13:51, 19 October 2017


Toehold switch for hsa-miR-15b-5p with luciferase

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 microRNA hsa-miR-15b-5p (the trigger RNA). The toehold switch is activated by hsa-miR-15b-5p and regulates production of Luciferase. The luminescence intensity of Luciferase is proportional to the number of toehold switches activated (as the more switches activated, the greater the amount of luciferase is produced), thus indicating the levels of hsa-miR-15b-5p present (as the more micro-RNA there is, the greater the number of switches activated). This part can therefore be used to quantify the levels of hsa-miR-15b-5p.

The transcripts produced by this part are toxic to E. coli. We recommend the use of a non-leaky promoter (for reference, we found BBa_K808000 was too leaky).

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 SapI.rc site found at 928