Part:BBa_K3365000

dCas9-ω

An RNA polymerase ω subunit is fused to the C-terminal of dCas9.

Usage and Biology

The dCas9 is a Cas9 nuclease mutant. Mutations D10A and H840A in the RucC and HNH domains, respectively, abolish cleavage but do not impair DNA binding. The dCas9 provides a simple and robust technology for gene repression and activation, and can target almost any DNA sequence aided by the sgRNA. The ω subunit can activate transcription by recruiting the RNAP holoenzyme. The fusion between dCas9 and ω subunit can activate gene transcription downstream of the protospacer. The CRISPR interference inhibits transcription by sterically blocking the RNA polymerase(RNAP).

Gene circuit of dCas9 with subnit Omega.png

Fig: Gene circuit

Working mode of dCas9 with subnit Omega.png

Fig:The figure above is shown how this part work in bacterial. The geneⅢ mentioned can be changed to any gene sequence depends on designed sgRNA.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1340
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1340
Illegal NheI site found at 1099
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1340
Illegal BamHI site found at 3378
Illegal BamHI site found at 4212
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1340
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1340
1000
COMPATIBLE WITH RFC[1000]