DNA

Part:BBa_K4438120

Designed by: Neha Adarsh   Group: iGEM22_IISER-Tirupati_India   (2022-10-11)

FASTmiR-222-D1_s

FASTmiR stands for Fluorescence Aptamer Sensor For Tracking miRNAs. It is an RNA-based sensor for in-vitro quantification of miRNAs. This part is the expression cassette for BBa_K4438100 with BBa_J64997 upstream and BBa_K731721 downstream along with some spacer sequences BBa_K3831027 (at 5’ end) and BBa_K4438136 (at 3’ end). FASTmiR-222-D1_s, after transcription, binds to miR-222, the transcription product of BBa_K4438150. The binding of these to RNAs will lead to the revelation of a binding site for DFHBI, a fluorophore, where it gets trapped and shows fluorescence.


Usage and Biology

This part can be used for the detection of miR-222, which proves to be a biomarker for many diseases. A promoter sequence added in the beginning enhances its usage in both in-vivo and in-vitro conditions. The OFF(A) and ON(A) structure for the sensor was generated as given: T--IISER-Tirupati India--miRNA 222 1.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 101
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 176
    Illegal SpeI site found at 101
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 101
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 101
  • 1000
    COMPATIBLE WITH RFC[1000]


Notes

The split position of the complementary sequence in the sensor is ACCCAGTAGCCA….GAGTGAGCT

References

Huang, K., Doyle, F., Wurz, Z. E., Tenenbaum, S. A., Hammond, R. K., Caplan, J. L., & Meyers, B. C. (2017). FASTmiR: an RNA-based sensor for in vitro quantification and live-cell localization of small RNAs. Nucleic acids research, 45(14), e130-e130

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