Difference between revisions of "Part:BBa K2206004"

Latest revision as of 23:05, 30 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 from 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 microRNA 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. Therefore, we recommend the use of an inducible non-leaky promoter to allow for amplification in E. coli. For reference, we found BBa_K808000 was a suitable replacement for BBa_J23111 when regulating a similar construct in E. coli that contained GFPmut3b in place of luciferase.

This part contains a strong RBS sequence.

NUPACK Structure Analysis

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

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 __NOTOC__
 <partinfo>BBa_K2206004 short</partinfo>
-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, resulting in translation of the 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. Luminescence intensity from Luciferase is proportional to the number of toehold switches activated, thus indicating the levels of hsa-mir-15b-5p present.
+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 from 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 microRNA 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. Therefore, we recommend the use of an inducible non-leaky promoter to allow for amplification in E. coli. For reference, we found BBa_K808000 was a suitable replacement for BBa_J23111 when regulating a similar construct in E. coli that contained GFPmut3b in place of luciferase.
+This part contains a strong RBS sequence.
+{{Template:CLSB-UK 17 Images 15p5p 1}}
 <!-- Add more about the biology of this part here