Difference between revisions of "Team:BostonU/Description"

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Project Design

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Abstract

While natural systems integrate diverse “digital” signals to precisely specify “analog” gene expression levels, synthetic systems thus far have focused on controlling expression in either a digital or an analog capacity. Our team sought to develop a “digitized-analog” expression system using CRISPR-dCas9, capable of specifying varied exogenous gene expression levels based on different signals. We first developed digital elements by pairing gRNAs with minimal operator promoters and using dCas9 to transactivate. We then created analog elements by multimerizing operator sites to obtain graded activation levels. Finally, we integrated our digital and analog elements into higher-order genetic logic circuits to achieve varying expression responses. We characterized and optimized our system in human cells, enabling synthetic biologists to better control transgene expression for important therapeutic applications..

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-<p style = "font-size:150%; padding:25px 150px 50px 150px; color:#0071A7;">Our project aims to integrate multiple digital environmental signals (signals that are either present or absent, no in between) to dictate the analog expression level of a certain gene. Different combinations of these digital signals will produce different intensities of gene expression. This is similar to many gene activation pathways found in nature, and yet there is no standardized, easy-to-use system to replicate these pathways. Being able to dynamically change the level of gene expression based on combinations of multiple signals would be invaluable to creating responsive, dynamic genetic devices. Therefore, our goal this summer is to create a system that can recognize multiple digital signals, and change the level of gene expression based on what combination of signals it is registering. This system would work with any gene of interest, allowing it to be used in diverse applications including immune therapy, recreating natural genetic signaling pathways, and control of toxic genes.</p>
+<p style = "font-size:150%; padding:25px 150px 50px 150px; color:#0071A7;">While natural systems integrate diverse “digital” signals to precisely specify “analog” gene expression levels, synthetic systems thus far have focused on controlling expression in either a digital or an analog capacity. Our team sought to develop a “digitized-analog” expression system using CRISPR-dCas9, capable of specifying varied exogenous gene expression levels based on different signals. We first developed digital elements by pairing gRNAs with minimal operator promoters and using dCas9 to transactivate. We then created analog elements by multimerizing operator sites to obtain graded activation levels. Finally, we integrated our digital and analog elements into higher-order genetic logic circuits to achieve varying expression responses. We characterized and optimized our system in human cells, enabling synthetic biologists to better control transgene expression for important therapeutic applications..</p>
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