Team:Penn/Project

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Project Description
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The code of life is more than a sequence of A’s, C’s, T’s and G’s; epigenetic modifications, such as DNA methylation, are powerful and heritable regulators of gene expression. Targeted methyltransferases are enzymes that catalyze sequence-specific methylation – the most useful tool for engineering the epigenome. With a synthetic biology approach, we developed an assay to test targeted methyltransferases without expensive, time-consuming traditional methods. Our modular single-plasmid system allows methyltransferases to be easily cloned and tested via inexpensive digestion assays, quickly measuring the existence and extent of targeted methylation. Additionally, our plasmid contains standardized primer-binding sites for methylation-sensitive sequencing, and our E. coli chassis effectively eliminated noise associated with methylation studies. We are using this assay to characterize our novel targeted methyltransferases, which could be used to study epigenetic modifications. In the future, synthetic biologists could embrace these tools to explore the next frontier in engineering biological systems: the epigenome.</center>
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                <p>The code of life is much more than a sequence of A's, G's, C's and T's;
 
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        a suite of epigenetic mechanisms, ranging from chromatin remodeling to non-coding RNAs,
 
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        affect gene expression and cellular function. <div class="figure-wrap" style="margin-left: 200px;"><img class="figure" src="https://googledrive.com/host/0B4ZBZOYYKBzEeG5XLTdURjc0aTA" "/>
 
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        <p> In particular, DNA methylation has been
 
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        shown to alter transcriptional activity in a powerful, heritable manner.  Abnormal methylation
 
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        patterns are associated with diseases including immunodeficiency syndromes, neurodevelopmental
 
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        disorders, and many types of cancer.  Comprehensive understanding and control of DNA methylation
 
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        could be invaluable to researchers studying these diseases.</p>
 
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        <p>Synthetic biologists and geneticists are accustomed to turning genes on and off at will,
 
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        but the tools don’t exist to easily manipulate epigenetic patterns.  We are developing a novel
 
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        fusion protein that enables site-specific methylation, which can repress promoter activity with
 
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        high precision.  <div class="figure-wrap" style="float: center;"><img src="https://googledrive.com/host/0B4ZBZOYYKBzEdmZMalozd0pkSjg" style="height: 75px; margin: 5px; float: left;"/>
 
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        <img src="https://googledrive.com/host/0B4ZBZOYYKBzEdlExOEgtVlZYYUU" style="height: 75px; float: left; margin: 5px;"/></div><p>In coming years, this fusion protein could become a powerful tool for epigenetics
 
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        researchers looking to perform on/off studies in the vein of classical genetics, as well as an
 
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        orthogonal mode of repressing constitutive promoters for bacterial synthetic biologists.
 
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        Eventually, it could even give clinical researchers the means to restore healthy methylation
 
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        levels in many insofar-untreatable epigenetic diseases.</p>
 
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Latest revision as of 03:50, 28 September 2013

modeling

Project Description


The code of life is more than a sequence of A’s, C’s, T’s and G’s; epigenetic modifications, such as DNA methylation, are powerful and heritable regulators of gene expression. Targeted methyltransferases are enzymes that catalyze sequence-specific methylation – the most useful tool for engineering the epigenome. With a synthetic biology approach, we developed an assay to test targeted methyltransferases without expensive, time-consuming traditional methods. Our modular single-plasmid system allows methyltransferases to be easily cloned and tested via inexpensive digestion assays, quickly measuring the existence and extent of targeted methylation. Additionally, our plasmid contains standardized primer-binding sites for methylation-sensitive sequencing, and our E. coli chassis effectively eliminated noise associated with methylation studies. We are using this assay to characterize our novel targeted methyltransferases, which could be used to study epigenetic modifications. In the future, synthetic biologists could embrace these tools to explore the next frontier in engineering biological systems: the epigenome.

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Retrieved from "http://2013.igem.org/Team:Penn/Project"