Composite

Part:BBa_K4409009:Design

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


igRNA (AAG+GAPDH 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 AAG present designed by previous groups is utilized to achieve a high sensitivity for the igRNA and Cas12 enzyme. The guide sequence targeting GAPDH dsDNA is chosen because GAPDH dsDNA is easily obtained in high quantity due to the abundant presence of GAPDH genes in cells and can be easily amplified. 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

This part is based on BBa_K2961003 for Cas12 handle with AAG present, BBA_K4409017 for the guide sequence, and BBa_K4409004 for the trigger-RNA sensor.

References