Difference between revisions of "Part:BBa K3365000"
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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). | 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). | ||
| + | [[File:Gene_circuit_of_dCas9_with_subnit_Omega.png]] | ||
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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. | ||
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Revision as of 08:55, 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).
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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1340
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1340
Illegal NheI site found at 1099 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1340
Illegal BamHI site found at 3378
Illegal BamHI site found at 4212 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1340
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1340
- 1000COMPATIBLE WITH RFC[1000]