RNA

Part:BBa_K2206013

Designed by: Sammy Lovat   Group: iGEM17_CLSB-UK   (2017-10-26)
Revision as of 10:22, 31 October 2017 by SLovat (Talk | contribs)

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Anti-miRNA for hsa-miR-15b-5p

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 sequence can be split into different RNA molecules. Colocalization of the split trigger parts results in a complete trigger RNA that is capable of activating the toehold switch. The formation of the duplex occurs through a highly specific binding step which results in single-base mismatch specificity.

The trigger RNA sequence for BBa_K2206012 is divided between hsa-miR-15b-5p and an anti-miRNA. This part is the anti-miRNA required for complete trigger RNA formation and activation of BBa_K2206012. This part contains two hybridisation domains: one that is complementary to the 3' end of hsa-miR-15b-5p and one which is complimentary to the 5' end of BBa_K2206012.

For more information, see the section on second series of toehold switches on the design page of CLSB-UK's 2017 wiki - http://2017.igem.org/Team:CLSB-UK/Design

NUPACK Structure Analysis

anti-miRNAmiRNAFree energy of secondary structure: -18.57 kcal/molMFE structure at 35.0°CACGU Free energy of secondary structure: -15.27 kcal/molMFE structure at 35.0°CACGUTrigger Binding SiteStart CodonRBS Free energy of secondary structure: -68.61 kcal/molMFE structure at 35.0°CACGUStart CodonTrigger Binding Siteanti-miRNAmiRNARBS

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
    COMPATIBLE WITH RFC[1000]


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Parameters
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