Part:BBa_K4409018

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

This is an RNA-interacting guide RNA (igRNA). It is used in a CRISPR-Cas12 system. It mainly comprises the following three parts. The first part (BBa_K2965036) is the Cas12 handle which combines with Cas12 protein. The second part (BBa_K4409001) is a guide sequence targeting a dsDNA sequence based on hsa_circ_0001785. The last part (BBa_K4409008) is a control sequence made up of VR1, loop and VR2 regions in sequence. VR1 and VR2 regions can specifically bind to the RNA trigger which is a unique sequence of hsa_circ_0001982. The loop binds to a portion of the guide sequence. When hsa_circ_0001982 is present in the CRISPR-Cas12 system, the control sequence binds with it and the igRNA is activated. The loop then unblocks the guide. The guide binds to a dsDNA with the hsa_circ_0001785 sequence and the PAM sequence, and activates the Cas12 protein, resulting in the cleavage of dsDNA and ssDNA in the system.

Graph 1 Figure Schematic representation of the inactive (A) and active(B,C) igRNA in a CRISPR-Cas12 system.

Sequence and Features

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]