Difference between revisions of "Part:BBa K3799000"

Latest revision as of 21:03, 21 October 2021

EiCsm6,A CRISPR system endoribonuclease

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. Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 314
Illegal XhoI site found at 367
Illegal XhoI site found at 1054
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K3799000 short</partinfo>
-This part encodes Csm6, a type III-A CRISPR-associated protein that degrades cyclic hexa-adenylate activator using both CARF and HEPN domains from Enterococcus italicus.cyclic hexa-AMP recognition by the CARF domain activates the Csm6 HEPN domains for collateral RNA degradation.It protects bacteria against transformation with plasmids containing DNA homologous to its spacer regions.
+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( [https://parts.igem.org/Part:BBa_K2323004 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.
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 ===Usage and Biology===