Difference between revisions of "Part:BBa K1150025"
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right" | {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right" | ||
| − | ! colspan="2" style="background:#FFBF00;"| | + | ! colspan="2" style="background:#FFBF00;"|CMV:HA-NLS-dCas9-Linker-PIF6-NLS:BGH |
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|'''Function''' | |'''Function''' | ||
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==Introduction== | ==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 | + | Our construct contains the standardized version of [https://parts.igem.org/Part:BBa_K1150000 dCas9] linked to [https://parts.igem.org/Part:BBa_K1150005 PIF6]. This device is one of two parts that you need to enable red-light induced gene targeting. Cotransfection of a PhyB+effector ([https://parts.igem.org/Part:BBa_K1150026 activation], [https://parts.igem.org/Part:BBa_K1150027 repression] or [https://parts.igem.org/Part:BBa_K1150028 epigenetical repression]) device and target RNA makes it possilbe 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 | + | The here used, mutated Cas9-version is human codon optimized and unable to cleave DNA. PIF6 is human codon-optimized, too. For more information and experimental data have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page]. |
| − | + | ||
| + | ==Data== | ||
| + | Data for this part are shown on the page of [https://parts.igem.org/Part:BBa_K1150026 Part II] | ||
<span class='h3bb'> | <span class='h3bb'> | ||
| + | |||
==Sequence and Features== | ==Sequence and Features== | ||
</span> | </span> | ||
Latest revision as of 01:53, 5 October 2013
uniCAS Red Light Switch Part I - Stimulator
| CMV:HA-NLS-dCas9-Linker-PIF6-NLS:BGH | |
|---|---|
| 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 (activation, repression or epigenetical repression) device and target RNA makes it possilbe 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 here used, mutated Cas9-version is human codon optimized and unable to cleave DNA. PIF6 is human codon-optimized, too. For more information and experimental data have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page].
Data
Data for this part are shown on the page of Part II
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]
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.