Difference between revisions of "Part:BBa K3365000"
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===Usage and Biology=== | ===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). | 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). | ||
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Revision as of 04:01, 24 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).
Sequence and Features
Assembly Compatibility:
- 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]