Difference between revisions of "Part:BBa K1150000"

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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1150000 short</partinfo>
 
<partinfo>BBa_K1150000 short</partinfo>
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
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! colspan="2" style="background:#FFBF00;"|dCas9
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|-
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|'''Function'''
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|Binding protein
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_25 RFC 25]
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|-
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|'''Backbone'''
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|pSB1C3<br>
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|-
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|'''Source'''
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|
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|-
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|'''Submitted by'''
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|Freiburg 2013
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|}
  
 
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 short RNA (called crRNA), which is in complex with Cas9, and the target DNA.<br>
 
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 short RNA (called crRNA), which is in complex with Cas9, 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 introduce mutations within the genome of several organisms by causing a double strand break. After the exchange of an aminoacid Cas9 was converted from a nuclease to a nickase, introducing only single strand breaks; and very recently converted to a enzymaticly inactive form, called dCas9, by another aminoacid exchange.<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 introduce mutations within the genome of several organisms by causing a double strand break. After the exchange of an aminoacid Cas9 was converted from a nuclease to a nickase, introducing only single strand breaks; and very recently converted to a enzymaticly inactive form, called dCas9, by another aminoacid exchange.<br>
 
The here available 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.
 
The here available 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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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'>
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==Sequence and Features==
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</span>
 
<partinfo>BBa_K1150000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1150000 SequenceAndFeatures</partinfo>
  

Revision as of 08:40, 24 September 2013

dCas9

dCas9
Function Binding protein
RFC standard RFC 25
Backbone pSB1C3
Source
Submitted by Freiburg 2013

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 short RNA (called crRNA), which is in complex with Cas9, 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 introduce mutations within the genome of several organisms by causing a double strand break. After the exchange of an aminoacid Cas9 was converted from a nuclease to a nickase, introducing only single strand breaks; and very recently converted to a enzymaticly inactive form, called dCas9, by another aminoacid exchange.
The here available 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]