Difference between revisions of "Part:BBa K1936001"
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[[File:Team_optoptosis_2016_lovbaxfull.png|750px|thumb|center|Fig.1 Activation of the blue light dependent OMM recruitment system.]]<html><br></html> | [[File:Team_optoptosis_2016_lovbaxfull.png|750px|thumb|center|Fig.1 Activation of the blue light dependent OMM recruitment system.]]<html><br></html> | ||
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+ | For our project we utilized the light dependent OMM recruitment as a potential method for cancer treatment. As shown in Fig.1 it recruits BAX protein which is fused to the ePDZ domain (see <html><a href="https://parts.igem.org/Part:BBa_K1936000">BBa_K1936000</a></html>). Once BAX is localized at the mitochondrial membrane it leads to formation of apoptotic channels. This allows us to kill abnormal cells with attainment of a very high level of spatiotemporal specificity compared to traditional cancer therapies (http://2016.igem.org/Team:Duesseldorf). | ||
===Biology=== | ===Biology=== |
Revision as of 17:10, 16 October 2016
Tom5-eGFP-LOV2
ALERT !!!Under construction !!
Usage
While LOV2 domain was previously described for optogenetical control of gene expression (BBa_K1742000) and as reporter gene (BBa_K660003), we took a new approach. The described fusion protein Tom5-eGFP-LOV2 is designed to recruit proteins of choice to the outer mitochondrial membrane (OMM) in engineered eukaryotic cells.
Tom5 is a part of the eukaryotic translocase of outer membrane protein complex and has a single membrane anchor unit that integrates into OMM. [1] We fused it to the N-terminal end of LOV2 domain from A.sativa. Upon exposure to blue light (λ=473nm) the LOV2 peptide undergoes a conformational change and exposes its C-terminal J-alpha-helix which is then able to bind a specifically engineered PDZ domain (BBa_K1470005). [2] The fused enhanced green fluorenscence protein (eGFP) allows the tracking of cellular localization by microscopy.
Our blue light dependent system can be used to recruit any protein of interest to the outer mitochondrial membrane by simply fusing it to an ePDZ domain.
For our project we utilized the light dependent OMM recruitment as a potential method for cancer treatment. As shown in Fig.1 it recruits BAX protein which is fused to the ePDZ domain (see BBa_K1936000). Once BAX is localized at the mitochondrial membrane it leads to formation of apoptotic channels. This allows us to kill abnormal cells with attainment of a very high level of spatiotemporal specificity compared to traditional cancer therapies (http://2016.igem.org/Team:Duesseldorf).
Biology
Characterization
References
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 33
Illegal BamHI site found at 712
Illegal XhoI site found at 515 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1226