Composite

Part:BBa_K4409018:Design

Designed by: Kexin Fei   Group: iGEM22_Worldshaper-HZ   (2022-09-20)


igRNA (CT+ hsa_circ_0001785 as guide+ hsa_circ_0001982 as trigger)


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]


Design Notes

The sequence of Cas12 handle with CT present designed by previous groups is utilized to achieve a high sensitivity for the igRNA and Cas12 enzyme. The guide sequence targeting a dsDNA with hsa_circ_0001785 sequence is used. This serves as a so-called double insurance. Only when both target is present will the fluorescence be produced. Thus, it greatly reduces the possibility of false positive to happen. The control sequence of the igRNA is designed so that it recognizes a unique sequence of hsa_circ_0001982 ranging across the back-splicing junction site. Therefore, the igRNA can specifically identify hsa_circ_0001982.


Source

The sequence that we designed is based on BBa_K2965036 for Cas12 handle with CT present, BBa_K4409001 for a guide sequence targeting a dsDNA sequence of hsa_circ_0001785, and BBa_K4409008 for detecting the trigger RNA of hsa_circ_0001982.

References