Difference between revisions of "Part:BBa K3365000"
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[[File:Action_mode_of_dCas9_with_subnit_Omega.png]]
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<b>Fig:</b>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.
 
<b>Fig:</b>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.

Revision as of 08:59, 27 October 2020


dCas9-ω

An RNAP ω subunit, RpoZ is fused to dCas9 C-terminal.

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


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]