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− | <br><p style = "font-size:150%; padding:5px 150px 15px 150px; color:#0071A7;">The BostonU 2016 iGEM team created Gemini, a design space that combines digital and analog expression systems to easily modulate exogenous gene expression levels in human cells. The system relies on three components: a genome-orthogonal gRNA recognizes a corresponding DNA operator sequence upstream of a minimal promoter, and it recruits dCas9-VPR to transactivate the output gene. We developed a set of mutually-orthogonal gRNAs to enable multiplexed gene regulation without cross-talk, and a set of gRNA-operator plasmids to achieve varied expression levels. Our submitted parts collection has 2 gRNA expression devices (<a href = "http://parts.igem.org/Part:BBa_K1875011" style = "color:blue;">BBa_K1875011</a>-<a href = "http://parts.igem.org/Part:BBa_K1875012" style = "color:blue;">BBa_K1875012</a>) and 7 gRNA-operator devices (<a href = "http://parts.igem.org/Part:BBa_K1875013" style = "color:blue;">BBa_K1875013</a>-<a href = "http://parts.igem.org/Part:BBa_K1875019" style = "color:blue;">BBa_K1875019</a>). We validated these parts using flow cytometry. We demonstrated “digital” expression when comparing output gene activation with or without gRNAs, and “analog” expression when comparing different gRNA-operator architectures (single, multimerized, and mutated sequences). </p> | + | <br> |
+ | <p style = "font-size:150%; padding:5px 150px 15px 150px; color:#0071A7;"> | ||
+ | The BostonU 2016 iGEM team created Gemini, a design space that combines digital and analog expression systems to easily modulate exogenous gene expression levels in human cells. The system relies on three components: a genome-orthogonal gRNA recognizes a corresponding DNA operator sequence upstream of a minimal promoter, and it recruits dCas9-VPR to transactivate the output gene. We developed a set of mutually-orthogonal gRNAs to enable multiplexed gene regulation without cross-talk, and a set of gRNA-operator plasmids to achieve varied expression levels. Our submitted parts collection has 2 gRNA expression devices ((<a href = "http://parts.igem.org/Part:BBa_K1875011" style = "color:blue;">BBa_K1875011</a>-<a href = "http://parts.igem.org/Part:BBa_K1875012" style = "color:blue;">BBa_K1875012</a>)) and 7 gRNA-operator devices ((<a href = "http://parts.igem.org/Part:BBa_K1875013" style = "color:blue;">BBa_K1875013</a>-<a href = "http://parts.igem.org/Part:BBa_K1875019" style = "color:blue;">BBa_K1875019</a>)). We validated these parts using flow cytometry. We demonstrated “digital” expression when comparing output gene activation with or without gRNAs, and “analog” expression when comparing different gRNA-operator architectures (single, multimerized, and mutated sequences). </p> | ||
<p style = "font-size:150%; padding:5px 150px 15px 150px; color:#0071A7;"> | <p style = "font-size:150%; padding:5px 150px 15px 150px; color:#0071A7;"> | ||
− | The team created pages for parts | + | The team created pages for parts BBa_K1875000 - BBa_K1875019 and submitted parts BBa_K1875011 - BBa_K1875019.</p> |
Revision as of 23:30, 18 October 2016
The BostonU 2016 iGEM team created Gemini, a design space that combines digital and analog expression systems to easily modulate exogenous gene expression levels in human cells. The system relies on three components: a genome-orthogonal gRNA recognizes a corresponding DNA operator sequence upstream of a minimal promoter, and it recruits dCas9-VPR to transactivate the output gene. We developed a set of mutually-orthogonal gRNAs to enable multiplexed gene regulation without cross-talk, and a set of gRNA-operator plasmids to achieve varied expression levels. Our submitted parts collection has 2 gRNA expression devices ((BBa_K1875011-BBa_K1875012)) and 7 gRNA-operator devices ((BBa_K1875013-BBa_K1875019)). We validated these parts using flow cytometry. We demonstrated “digital” expression when comparing output gene activation with or without gRNAs, and “analog” expression when comparing different gRNA-operator architectures (single, multimerized, and mutated sequences).
The team created pages for parts BBa_K1875000 - BBa_K1875019 and submitted parts BBa_K1875011 - BBa_K1875019.