Composite

Part:BBa_K3110051

Designed by: Amartya Pal   Group: iGEM19_IISER_Tirupati   (2019-10-16)
Revision as of 14:53, 20 October 2019 by Bhabesh (Talk | contribs)


lldRO1-J23117-lldRO2 Strong RBS YebF-IL12

lldRO1 and lldRO2 are the operator regions which bind to the regulator element lldR and inhibit transcription. Upon binding of L-Lactate to lldR, however, this transcriptional suppression is lost and instead lldR complex with L-Lactate remains bound to lldRO1 acting as a transcriptional activator. By using RBS of different strengths we are trying to find out the optimum construct which will give output (in this case the output is YebF-IL12) that can efficiently discriminate cancer vs non-cancer micro-environments and will lead to secretion of IL12 (natural immuno-modulator) and eventually reduction of tumour size.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 78
    Illegal NheI site found at 101
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology<h1> The regulatory element lldR binds to the lldRO1 and lldRO2 (the operator regions) and inhibit transcription. J23117 is a promoter intercalated between the operators. lldR dimer represses the transcription possibly by forming a DNA loop which doesn’t allow the RNA polymerase to bind the promoter. Upon binding of L-Lactate to LldR, however, this transcriptional suppression is lost and instead lldR complex with L-Lactate remains bound to LldRO1 acting as a transcriptional activator. Interleukin-12. IL-12 is an anti-inflammatory cytokine, involved in the regulation of cell-mediated immune responses. Through a set of complex signaling cascade IL-12 activates our immune cells to reduce tumor growth. It is under the control of lactate sensitive O1PO2 to promote its production only when the cells encounter a thresold concentration of lactic acid. It is conjugated to YebF that mediates its secretion outside the cells into the cancer microenvironment. It also has a flag tag attached downstream to YebF so that its secretion outside the cells can be detected using anti-Flag antibodies.

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