Types | DnaRegion
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Roles | CDS
Coding
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Sequences | BBa_K1218013_sequence (Version 1)
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Description
CRISPR-Cas is a bacterial immune system that remembers and targets foreign viral DNA by storing DNA sequences, or spacers, between clustered regularly interspaced short palindromic repeats (CRISPRs). RNA transcripts of the spacers are then used to sense homologous DNA, which is cleaved by CRISPR-associated (Cas) proteins.
This part codes for the tracrRNA, a mutated Cas9 protein, and minimal CRISPR array of a type II CRISPR-Cas system. The CRISPR array includes two CRISPR repeats separated by a spacer with two BsaI sites. Digestion with BsaI allows for insertion of a new spacer, thus changing the sequence targeted by Cas9. However, because the Cas9 protein is mutated or "dead", the targeted gene is merely repressed by dCas9.
Notes
Mutagenesis to remove internal EcoRI site
Source
CRISPR-Cas system from streptococcus pyogenes; cloned from plasmids obtained from Bikard et al.