Difference between revisions of "Part:BBa K1150000"

Revision as of 14:54, 21 September 2013

dCas9

Cas9 is the main protein of the CRISPR-Cas system of Streptococcus pyogenes, which is categorized as CRISPR system type II. Like all other CRISPR systems it protects bacteria (and archaea) from phages by recognizing and cleaving of the invading phage DNA. This recognition is based on Watson Crick base pairing between a complexed short RNA (called crRNA) and the target DNA.
Because of the ability to recognize almost every DNA sequenz, Cas9 became of interest for research concerning DNA targeting. At first it was used together with the crRNA and a tracrRNA, which is required to form the protein RNA complex, to intoduce mutations within the genome of several organisms by causing a double strand break. After the exchange of a aminoacid Cas9 was converted from a nuclease to a nickase, introducing only single strand breaks; and very recently converted to a enzymatic inactive form, called dCas9, by another aminoacid exchange.
The here avaliable dCas9 is codon optimized for human cell lines and standardized (RFC 25). It can be used as a DNA binding protein, that can be fused with different effectors in order to regulate gene expression.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 248
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K1150000 short</partinfo>
-The human codon optimized Cas9 from the organism Streptococcus pyogenes was standardized (RFC 25). Additionally we mutated the Nickase function. In the end we engineered a DNA binding protein, that can be fused with different effectors in order to regulate gene expression.
+Cas9 is the main protein of the CRISPR-Cas system of <i>Streptococcus pyogenes</i>, which is categorized as CRISPR system type II. Like all other CRISPR systems it protects bacteria (and archaea) from phages by recognizing and cleaving of the invading phage DNA. This recognition is based on Watson Crick base pairing between a complexed short RNA (called crRNA) and the target DNA.<br>
+Because of the ability to recognize almost every DNA sequenz, Cas9 became of interest for research concerning DNA targeting. At first it was used together with the crRNA and a tracrRNA, which is required to form the protein RNA complex, to intoduce mutations within the genome of several organisms by causing a double strand break. After the exchange of a aminoacid Cas9 was converted from a nuclease to a nickase, introducing only single strand breaks; and very recently converted to a enzymatic inactive form, called dCas9, by another aminoacid exchange.<br>
+The here avaliable dCas9 is codon optimized for human cell lines and standardized (RFC 25). It can be used as a DNA binding protein, that can be fused with different effectors in order to regulate gene expression.
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