Composite
Part:BBa_K4409010:Design
Designed by: Kexin Fei Group: iGEM22_Worldshaper-HZ (2022-09-20)
igRNA (CT+GAPDH as guide+ hsa_circ_0001982 as trigger)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The sequence of Cas12a handle with CT present 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_K4409000 for Cas12 handle with CT present, BBA_K4409017 for the guide sequence, and BBa_K4409004 for the trigger-RNA sensor.