Team:Debrecen-Hungary/project

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=Abstract=
=Abstract=
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Animal synthetic biology has huge potential, yet it is still in need of more diverse molecular tools for defined gene regulation.
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Nuclear receptors are a conserved family of proteins responsible for sensing lipids; they may be viewed as lipid activated transcription factors.
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We have successfully developed a kit with a variety of lipid responsive domains (from H.sapines, D.melanogaster and C.elegans) for the rational construction of synthetic transcription factors. The domains respond only to predefined lipids and selectively activate predetermined gene expression.
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To characterize theses domains, we used standardized protocols for comparable measurements. In vivo gene expression was measured as a function of ligand concentration using luciferase activity.
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The potential for these tools is immense; e.g. from the ultra sensitive detection of lipid contaminants in the environment to the opportunity of titration specific gene expression canges in patients undergoing gene therapy.

Revision as of 10:39, 19 October 2010



Abstract

Animal synthetic biology has huge potential, yet it is still in need of more diverse molecular tools for defined gene regulation. Nuclear receptors are a conserved family of proteins responsible for sensing lipids; they may be viewed as lipid activated transcription factors. We have successfully developed a kit with a variety of lipid responsive domains (from H.sapines, D.melanogaster and C.elegans) for the rational construction of synthetic transcription factors. The domains respond only to predefined lipids and selectively activate predetermined gene expression. To characterize theses domains, we used standardized protocols for comparable measurements. In vivo gene expression was measured as a function of ligand concentration using luciferase activity. The potential for these tools is immense; e.g. from the ultra sensitive detection of lipid contaminants in the environment to the opportunity of titration specific gene expression canges in patients undergoing gene therapy.






Our team has decided to focus efforts at making a nuclear hormone receptor kit, for use in eukaryotic cells. Our attention was drawn to this class of receptors due to their key physiological contribution in the endocrine system, cellular differentiation pathways, paracrine signaling and more.

 Upon ligand binding, nuclear hormone receptors undergo a change in conformation. They then translocate to the nucleus where they bind to specific DNA elements. The binding ultimately leads to defined changes in gene expression (both activation and repression). They may be viewed as special transcription factors which can be activated by extracellular cues.

The kit will include various basic parts of which a user might be able to create his own ligand activated transcription factor (a composite part). Our primary goal is thorough characterization of each basic part. Our methods and results as well as secondary goals will be addressed at a later time.

We hope that our biobrick part contribution may be used in a variety of medicinal applications as well as environmental sustainability projects or even other fields.












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