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Team:BostonU/Future
From 2014.igem.org
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We are in the process of gathering <a href="https://2014.igem.org/Team:BostonU/Data">flow cytometry data</a> for our basic parts library to complete Phase I of the Chimera workflow. | We are in the process of gathering <a href="https://2014.igem.org/Team:BostonU/Data">flow cytometry data</a> for our basic parts library to complete Phase I of the Chimera workflow. | ||
| - | We have begun carrying out <a href="https://2014.igem.org/Team:BostonU/Multiplexing">multiplexing reactions</a> as part of Phase II. We plan to gather data for the range of functionality of our transcriptional unit variants and begin assembling our priority encoder from its constitutive transcriptional units. Before building the full priority encoder, we will finish testing the new repressor genes, promoters, tandem promoters, and fusion proteins. We will prioritize pBad-pBetI and pLmrA-pSrpR. Once we confirmed the function of these parts and the correct regulatory arcs between parts, we will continue on to building the priority encoder. Functionality of the priority encoder via flow cytometry verification will mark the completion of Phase III and the Chimera workflow.<br><br> | + | We have begun carrying out <a href="https://2014.igem.org/Team:BostonU/Multiplexing">multiplexing reactions</a> as part of Phase II. We plan to gather data for the range of functionality of our transcriptional unit variants and begin assembling our priority encoder from its constitutive transcriptional units. Before building the full priority encoder, we will finish testing the new repressor genes, promoters, tandem promoters, and fusion proteins. We will prioritize the tandem promoters: pBad-pBetI and pLmrA-pSrpR, fusion proteins: BetI-GFP and LmrA-GFP, and the origin of replication p15A. Once we confirmed the function of these parts and the correct regulatory arcs between parts, we will continue on to building the priority encoder. Functionality of the priority encoder via flow cytometry verification will mark the completion of Phase III and the Chimera workflow.<br><br> |
<h2>Refine Chimera</h2><br> | <h2>Refine Chimera</h2><br> | ||
The testing of our workflow by building the priority encoder is intended to demonstrate the ability of Chimera's methodology and software tools to guide a predictive assembly of a complex genetic regulatory network. We will consider our documented difficulties in cloning, testing, and interpreted data t to refine our workflow and, most importantly, define concrete metrics that must be met at each phase of the workflow. These metrics must be adaptable to any user's target device, and must therefore allow for flexibility in their definition. For example, our version of the workflow currently specifies that we screen 10 colonies per plate if we have multiplexed 5 different basic parts to allow testing of a higher range of TU variety without excessive screening. This metric would be different for protocols that measure TU variety without multiplexing in quantity, but would still be present to ensure that an appropriate variety of gene expression is being observed.<br><br> | The testing of our workflow by building the priority encoder is intended to demonstrate the ability of Chimera's methodology and software tools to guide a predictive assembly of a complex genetic regulatory network. We will consider our documented difficulties in cloning, testing, and interpreted data t to refine our workflow and, most importantly, define concrete metrics that must be met at each phase of the workflow. These metrics must be adaptable to any user's target device, and must therefore allow for flexibility in their definition. For example, our version of the workflow currently specifies that we screen 10 colonies per plate if we have multiplexed 5 different basic parts to allow testing of a higher range of TU variety without excessive screening. This metric would be different for protocols that measure TU variety without multiplexing in quantity, but would still be present to ensure that an appropriate variety of gene expression is being observed.<br><br> | ||
Revision as of 03:19, 16 October 2014
Complete priority encoderWe are in the process of gathering flow cytometry data for our basic parts library to complete Phase I of the Chimera workflow. We have begun carrying out multiplexing reactions as part of Phase II. We plan to gather data for the range of functionality of our transcriptional unit variants and begin assembling our priority encoder from its constitutive transcriptional units. Before building the full priority encoder, we will finish testing the new repressor genes, promoters, tandem promoters, and fusion proteins. We will prioritize the tandem promoters: pBad-pBetI and pLmrA-pSrpR, fusion proteins: BetI-GFP and LmrA-GFP, and the origin of replication p15A. Once we confirmed the function of these parts and the correct regulatory arcs between parts, we will continue on to building the priority encoder. Functionality of the priority encoder via flow cytometry verification will mark the completion of Phase III and the Chimera workflow. Refine ChimeraThe testing of our workflow by building the priority encoder is intended to demonstrate the ability of Chimera's methodology and software tools to guide a predictive assembly of a complex genetic regulatory network. We will consider our documented difficulties in cloning, testing, and interpreted data t to refine our workflow and, most importantly, define concrete metrics that must be met at each phase of the workflow. These metrics must be adaptable to any user's target device, and must therefore allow for flexibility in their definition. For example, our version of the workflow currently specifies that we screen 10 colonies per plate if we have multiplexed 5 different basic parts to allow testing of a higher range of TU variety without excessive screening. This metric would be different for protocols that measure TU variety without multiplexing in quantity, but would still be present to ensure that an appropriate variety of gene expression is being observed. Push Chimera to the limitThe software tools we employ have the capability of allowing for construction of regulatory networks with greater complexity that our priority encoder. We aim to use Chimera to construct a novel device with nonlinear behavior, which will involve controlled protein degradation and a more complex testing scheme. |  |
