Part:BBa_K3799026
Type III-A CRISPR-Cas effector ribonuclease---Csm6
Csm6 is a non-specific endoribonuclease that cleaves single-stranded RNA upon activation by cyclic adenylates or linear adenine homopolymers terminated with a 2'3'-cyclic phosphate.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
Description - EiCsm6 is obtained from Enterococcus italicus, and belongs to the type III CRISPR systems. Csm6 is mainly activated by cyclic oligoadenylate second messengers by binding to their CARF domain. This activates the HEPN domain of the Csm6 for collateral RNA degradation. The CARF domain then degrades the cyclic adenylate activator (cA6) which functions as an off switch to regulate the activity of the CRISPR enzyme. These mechanisms facilitate rapid invader clearance and ensures proper termination to limit self toxicity and suicide of the bacteria.
EiCsm6 is coupled with the enzyme LwaCas13a(BBa_K2323004) to create the detector system SHERLOCKv2. The overhangs that are produced by the cleavage of RNA by Cas13a produces 2’-3’ cyclic phosphate hex adenylate residues that activate Csm6. This activation results in the collateral cleavage of RNA by both Cas13a and Csm6.
Cloning and characterization
Cloning
| None |