Difference between revisions of "Part:BBa K1150025"
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|'''Function''' | |'''Function''' | ||
| − | | | + | |dCas9-protein linked to PIF6 |
induced gene expression control | induced gene expression control | ||
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=Introduction= | =Introduction= | ||
| − | Our construct contains the standardized version of | + | Our construct contains the standardized version of dCas9 linked to PIF6. This device is one of two parts that you need to enable red-light induced gene targeting. Cotransfection of a PhyB+effector device, and target RNA allows you to recruit any desired effector to your desired gene locus. Illumination with red light (660nm) will start the interaction and illumination with far red light (740nm) will stop the interaction. |
| − | The Cas9-version used here is human codon optimized and the mutated version | + | The Cas9-version used here is human codon optimized and the mutated dCas9-version is unable to cleave DNA. PIF6 is human codon-optimized. |
=References:= | =References:= | ||
Muller, K.; Engesser, R.; Metzger, S.; Schulz, S.; Kampf, M. M.; Busacker, M. et al. ((Nucleid Acid Research; 2013): A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells. | Muller, K.; Engesser, R.; Metzger, S.; Schulz, S.; Kampf, M. M.; Busacker, M. et al. ((Nucleid Acid Research; 2013): A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells. | ||
Revision as of 23:34, 3 October 2013
uniCAS Red Light Switch Part I - Stimulator
| uniCAS red Light Switch Part I - Stimulator | |
|---|---|
| Function | dCas9-protein linked to PIF6
induced gene expression control |
| Use in | Mammalians |
| RFC standard | RFC 25 |
| Backbone | pSB1C3 |
| Submitted by | [http://2013.igem.org/Team:Freiburg Freiburg 2013] |
Introduction
Our construct contains the standardized version of dCas9 linked to PIF6. This device is one of two parts that you need to enable red-light induced gene targeting. Cotransfection of a PhyB+effector device, and target RNA allows you to recruit any desired effector to your desired gene locus. Illumination with red light (660nm) will start the interaction and illumination with far red light (740nm) will stop the interaction. The Cas9-version used here is human codon optimized and the mutated dCas9-version is unable to cleave DNA. PIF6 is human codon-optimized.
References:
Muller, K.; Engesser, R.; Metzger, S.; Schulz, S.; Kampf, M. M.; Busacker, M. et al. ((Nucleid Acid Research; 2013): A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 576
Illegal BglII site found at 900 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]