Difference between revisions of "Part:BBa K1150018"
| (2 intermediate revisions by the same user not shown) | |||
| Line 30: | Line 30: | ||
| − | Cas9 is a protein that binds DNA via an protein-RNA-DNA interaction. This version of Cas9 is working as a carrier for effectors that want to be brought to DNA in a sequence specific manner. The native nikase activity of the protein has been mutated so that it allows DNA binding without cutting. The sequence is human codon optimized. | + | Cas9 is a protein that binds DNA via an protein-RNA-DNA interaction. [https://parts.igem.org/Part:BBa_K1150000 This version of Cas9] (dCas9) is working as a carrier for effectors that want to be brought to DNA in a sequence specific manner. The native nikase activity of the protein has been mutated so that it allows DNA binding without cutting. The sequence is human codon optimized. |
| − | The protein is part of team Freiburg's (2013) the uniCAS toolkit. | + | The protein is part of team Freiburg's (2013) the [http://2013.igem.org/Team:Freiburg uniCAS toolkit]. |
| − | The usage of the medium strong [ | + | The usage of the medium strong [https://parts.igem.org/Part:BBa_K1150011 SV40 promoter] optimizes this device for means were a strong expression is suboptimal. If a strong expression is desired check our [https://parts.igem.org/Part:BBa_K1150017 device with CMV promoter]. |
==Usage and Biology== | ==Usage and Biology== | ||
| − | The | + | The dCas9 protein is able to target several loci at once as it interacts with small RNAs to build up a complex that will interact with complementary DNA strands. Its origin is the adaptive immune system of <i>Streptococcus pyogenes </i> called CRISPR. Hijacking this system leads to a whole new approach for multiple gene targeting. |
The iGEM team Freiburg 2013 combined effectors with this protein to create a toolkit, that is able to activate or repress specific and multiple target loci. | The iGEM team Freiburg 2013 combined effectors with this protein to create a toolkit, that is able to activate or repress specific and multiple target loci. | ||
Latest revision as of 16:53, 4 October 2013
Cas9 with SV40 promoter
| pSV40:HA-NLS-dCas9-G9a-NLS:tBGH | |
|---|---|
| Function | DNA binding protein |
| Use in | Mammalian cells |
| RFC standard | RFC 25 |
| Backbone | pSB1C3 |
| Organism | Streptococcus pyogenes |
| Source | Feng Zhang, Addgene |
| Submitted by | [http://2013.igem.org/Team:Freiburg Freiburg 2013] |
Cas9 is a protein that binds DNA via an protein-RNA-DNA interaction. This version of Cas9 (dCas9) is working as a carrier for effectors that want to be brought to DNA in a sequence specific manner. The native nikase activity of the protein has been mutated so that it allows DNA binding without cutting. The sequence is human codon optimized. The protein is part of team Freiburg's (2013) the [http://2013.igem.org/Team:Freiburg uniCAS toolkit].
The usage of the medium strong SV40 promoter optimizes this device for means were a strong expression is suboptimal. If a strong expression is desired check our device with CMV promoter.
Usage and Biology
The dCas9 protein is able to target several loci at once as it interacts with small RNAs to build up a complex that will interact with complementary DNA strands. Its origin is the adaptive immune system of Streptococcus pyogenes called CRISPR. Hijacking this system leads to a whole new approach for multiple gene targeting.
The iGEM team Freiburg 2013 combined effectors with this protein to create a toolkit, that is able to activate or repress specific and multiple target loci.
This approach offers new possibilities for fundamental epigenetic research, tissue engineering and cancer research.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 664
Illegal BglII site found at 5139 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]