Difference between revisions of "Team:Bordeaux/Description"

 
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<h1>Description</h1>
 
<h1>Description</h1>
  
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<p>This year, IGEM bordeaux aims to study the alternative splicing of the unc-60 gene in <i>C. elegans</i>. This gene can be spliced differently, leading to the synthesis of different proteins. The alternative splicing of Unc-60 depends on the precense of two proteins, Sup12 and Asd2. The proteins are expressed differently in different tissu of the organism. </p>
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<p>In this project, we are trying to control the alternative splicing of unc-60. To do so, we want to control the expression of one of the protein important for the splicing : Sup12. Thus, we will be able to choose which isoform of the protein encoded by unc-60 will be produced.  
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On the one hand, we want to make a photo-inductible system using Neurospora carassa's White-Collar system. The splicing protein will be under control of the Frq promoter, regulated by the White-Collar complex (expressed in the modified C.elegans). The White-Collar complex will fix the Frq promoter in response to light stimulation, allowing the expression of Sup12.
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On the other hand, we aim to use another inductible system called the Q system (from Neurospora crassa). This system responds to the presence of quinic acid and could allow a more specific control of the splicing protein Sup12. The protein will be under the control of the QUAS promoter. this promoter can be recognise by the protein QF, initiating the transcription. But the repressive protein QS inhibits this activation of transcription. This two protein, QF and QS will be expressed in specific tissus. By adding quinic acid in the growth medium, we should be able to prevent the action of QS and thus allow the expression of Sup12.
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We also aim to create a microfluidic system to observe the results of these genetic modifications on C. elegans.</p>
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This year, iGEM bordeaux 2017 aims to study the alternative splicing of the unc-60 gene in <i>C. elegans</i>. This gene can be spliced differently and two isoforms can be generated : UNC-60A, a non muscular isoform and UNC-60B, a muscular isoform. Thus, identity of cell depends on these two proteins and that is why our project focuses on the splicing of unc-60 which generate the muscular isoform. Our goal is to control this splicing by light activation through a mechanism known in <i>Neurospora crassa</i> (Fungi) or, alternatively, by chemical activation also known in <i>N. crassa</i>.</p>
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The alternative splicing of this gene is regulated by the SUP-12's RRM (RNA Recognition Motif) domain, a region of the protein which binds to the unc-60 pre-RNA, to induce, with ASD-2, the muscular isoform. Thus the cell becomes a muscular cell. The "Stargate WCC" project aims to control alternative splicing of unc-60 by SUP-12 protein. For this purpose, our team uses a photo-inducible system, the White Collar system which exists naturally in <i>N. crassa</i> in which it regulates the circadian rhythm according to the light level. Indeed, two proteins, White Collar 1 and 2, fit together in order to form a complex : the White Collar Complex or WCC, activated by a conformational change in response to light. Then this WCC modified and activated binds to circadian genes promoters in the aim of inducing FRQ proteins. In the worm, the WCC will be implied into neural cell to activate SUP-12 protein, regulate the alternative splicing of unc-60 and generate a muscle cell instead of a neural cell.</p>
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In conclusion, the "Stargate WCC" project is intended to control alternative splicing via different regulating mechanisms involving a logical gate that inspired us the project name. For this purpose, our team decided to focus on gene expression in muscular cell in order to control cell differentiation. This experimental procedure strives to give necessary tools to lead then some biomedical research linked to this phenomenon.</p>
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<h5>References</h5>
 
<h5>References</h5>
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<cite style=" color: #E0E0E0">Ohno G, Ono K, Togo M, Watanabe Y, Ono S, et al. (2012) Muscle-Specific Splicing Factors ASD-2 and SUP-12 Cooperatively Switch Alternative Pre-mRNA Processing Patterns of the ADF/Cofilin Gene in Caenorhabditis elegans. PLOS Genetics 8(10): e1002991. https://doi.org/10.1371/journal.pgen.1002991</cite>
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<cite style="color:#E0E0E0">Anyanful A, Ono K, Johnsen RC, et al. The RNA-binding protein SUP-12 controls muscle-specific splicing of the ADF/cofilin pre-mRNA in C. elegans. The Journal of Cell Biology. 2004;167(4):639-647. doi:10.1083/jcb.200407085.</cite>
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<cite style="color:#E0E0E0">Ballario P, Vittorioso P, Magrelli A, Talora C, Cabibbo A, Macino G. White collar-1, a central regulator of blue light responses in Neurospora, is a zinc finger protein. The EMBO Journal. 1996;15(7):1650-1657.</cite>
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<cite style="color:#E0E0E0">San-Miguel A, Lu H. Microfluidics as a tool for C. elegans research. In: WormBook: The Online Review of C. elegans Biology [Internet]. Pasadena (CA): WormBook; 2005-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK174829/</cite>
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<h1 style="text-align:center;color:#d8b700">How to find us ?</h1>
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<p style="font-size:1.5em; text-align:center; color:#E0E0E0">Feel free to email us to provide some feedback on our project, have some information on the team and our work, or to just say hello !</p>
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<h5 style="float:left ; color:white">Mail:
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<h5 style="text-align:right ; color:white"><i>Copyright &#169; iGEM Bordeaux 2017</i></h5>
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Latest revision as of 02:31, 2 November 2017

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