Types | DnaRegion
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Roles | CDS
Coding
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Sequences | BBa_K1723000_sequence (Version 1)
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Description
This part codes for a enzymatically inactive Cas9 protein (dCas9) that is fused to rpoZ, the omega subunit of E.Coli RNA Polymerase (RNAP).
Cas9 (CRISPR associated protein 9) is an RNA-guided DNA endonuclease that targets and cleaves any DNA sequence complementary to its guide RNA (gRNA). Catalytically ???dead??? Cas9 (dCas9) lack the ability to cleave DNA. When fused to a RNAP recruiting element (in this case the omega subunit of RNAP), chimeric dCas9 can act as a programmable transcription activator. dCas9 may also act as a DNA transcription inhibitor: depending on its gRNA-determined binding site, it has been shown in yeasts to sterically hinder RNAP recruitment to promoter sequences.
Notes
The original dCas9-ω given to us by the Bikard lab contains an EcoRI restriction site. We performed a site-directed mutagenesis to remove it.
Source
A plasmid containing dCas9_ω (pdCas9) was given to our iGEM team by David Bikard.