Plasmid

Part:BBa_K1391026

Designed by: Jing Wei "Raymond" Liu   Group: iGEM14_MIT   (2014-10-17)

High sensor: L7Ae_miR-let-7f

Our completed high sensor for detection of miR-let-7f, which is upregulated in Alzheimer's disease. Expresses L7ae which represses further downstream output when miR-let-7f concentration is high.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal NotI site found at 402
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal BglII site found at 1694
    Illegal BglII site found at 2980
    Illegal XhoI site found at 394
    Illegal XhoI site found at 2093
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal NgoMIV site found at 645
    Illegal NgoMIV site found at 926
    Illegal NgoMIV site found at 1828
    Illegal AgeI site found at 774
    Illegal AgeI site found at 1206
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2769
    Illegal BsaI.rc site found at 2881


A



B

C

High Sensor let-7f in the presence and absence of siRNA-7f in HEK293 cells. Using red as our transfection marker and green as our output, we expected that the sensor, when exposed to the inhibiting siRNA, would show an increase in GFP output. The cytometry results in figure (A) shows that there is no difference between the 'on' and 'off' state. This conclusion is reinforced by the microscopy in (B) and (C). On the left the sensor is in a low miRNA environment, and on the right the sensor is in a high miRNA environment.

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