Part:BBa_K1150018
| pSV40:HA-NLS-dCas9-G9a-NLS:tBGH | |
|---|---|
| Function | DNA binding protein fused to a methyl histone transferase |
| Use in | Mammalian cells |
| RFC standard | RFC 25 |
| Backbone | pSB1C3 |
| Organism | Streptococcus pyogenes, Mus musculus |
| Source | Feng Zhang, Addgene Albert Jeltsch, University of Stuttgart |
| Submitted by | [http://2013.igem.org/Team:Freiburg Freiburg 2013] |
Cas9 with SV40 promoter
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. The protein is part of team Freiburgs (2013) the 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 uniCAS Histone Modificator device with CMV promoter [[1]].
Usage and Biology
The dCAS9 protein is able to be targeted to 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.
References
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]
| None |