Team:Valencia UPV/Project/modules/biosafety

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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a>Biosafety</a> </h3></p><br/>
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<p>In order to make the Sexy Plant a safe plant, we developed a biosafety module which will be available for teams working with plants in future iGEM editions. The aim of the biosafety module is to avoid the dispersion of the plant’s genetic material in the environment. Two strategies were combined into the same module to make our plant safe: identity preservation and male sterility.</p><br/><br/>
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<div align="center"><span class="coda"><roja>B</roja>iosafety</span> </div><br/><br/>
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<p>Identity preservation enables an easy identification of the genetically modified organism. A chromoprotein, which provides a differential pigmentation to the plant, is incorporated in the module. The implementation of this chromoprotein to our system is the result of the collaboration with NRP-UEA-Norwich, and it is also used in their project “Green Canary”.</p><br/><br/>
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<p>"With great powers comes great responsibility." – <span class="italic">Benjamin Parker (Uncle Ben)</span>.</p><br/><br/>
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<p>Male sterility makes impossible the dispersion of genetic material using pollen as the vehicle. In order to achieve this dispersion restriction, the gene coding for barnase (a ribonuclease from Bacillus amyloliquefaciens) under the regulation of the tapetum-specific promoter TA29[1] is incorporated in our system. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8] . This strategy was chosen because it had been previously used in our laboratory with satisfactory results [9]</p><br/><br/>
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<p>We know the importance of keeping genetically modified organisms under control. Ideally, modified genetic material should be unable to spread and organisms containing it should be easily distinguishable from wild type. Keeping this in mind, we created a biosafety module to be used in plants which will be available for future iGEM teams working with plants. Our biosafety module combines two biosafety strategies: <span class="black-bold">identity preservation</span> and <span class="black-bold">male sterility</span>.</p><br/><br/>
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<p><span class="black-bold">Identity preservation</span> enables an easy identification of the genetically modified organism. We expressed SlANT1 and SlJAF13 genes from tomato, which code for transcription factors that activate synthesis of <span class="black-bold">anthocyanines </span> [1], coloured compounds found in plants. The introduction of this element in the biosafety module generates differentially coloured plants (Figures 1 and 2).  <a class="normal-link-page" target="_blank" href="https://2014.igem.org/Team:NRP-UEA-Norwich">NRP-UEA-Norwich</a> , and it is also used in their project “<span class="italic">Green Canary</span>”.</p><br/><br/>
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<div align="center"><img height= "170px;" style="margin-right: 30px;" src="https://static.igem.org/mediawiki/2014/b/b8/VUPV_Rosea.png" alt="N benthamiana Rosea" title=N benthamiana Rosea""></img><img width="250px" style="margin-left: 30px;" src="https://static.igem.org/mediawiki/2014/5/57/VUPVhoja.png" alt="Dark leaf" title="Dark leaf"></img><br/><br/>
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<p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>.Anthocyanin accumulation in a <i>N benthamiana</i> plant.                     
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      <p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><b>Figure 2</b>. Dark purple leaf with induced anthocyanin accumulation.  </p><br/>
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<p><span class="black-bold">Male sterility</span> makes impossible the dispersion of genetic material using pollen as the vehicle or by seeds result of self-pollination. In order to achieve this dispersion restriction, we integrated the active peptide of <span class="black-bold">barnase</span>, a RNAse from <span class="italic">Bacillus amyloliquefaciens</span>, (Biobricks accession code BBa_I716211) under the regulation of the tapetum-specific promoter TA29 [1]. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8]. We chose this strategy because it had been previously used in our laboratory with satisfactory results [9].</p><br/>
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<p>Our Biosafety module is our major present for future iGEM teams. It can be applied to a broad variety of new systems in future projects. We are certain that future iGEMers will realize how important biosafety is and they will incorporate our biosafety module in their systems.</p>
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<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>&larr; Go to Methodology</strong></a>
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<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules"><strong>Go to Modules</strong></a>
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<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/ideal_plant"><strong>Go to Future Perspectives &rarr;</strong></a></div></br></br></br>
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<br/><br/><p align="center"><strong>References</strong></p><br/>
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<div style="position: relative; left: 3%; width: 96%;"><ol>
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<li>Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737-741.</li>
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<li>Cho HJ, Kim S, Kim M, Kim BD (2001) Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells 11: 326-333.</li>
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<li>Sa G, Mi M, He-Chun Y, Guo-Feng L (2002) Anther-specific expression of ipt gene in transgenic tobacco and its effect on plant development. Transgenic Res 11: 269-278.</li>
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<li>Shaya F, Gaiduk S, Keren I, Shevtsov S, Zemah H, et al. (2012) Expression of mitochondrial gene fragments within the tapetum induce male sterility by limiting the biogenesis of the respiratory machinery in transgenic tobacco. J Integr Plant Biol 54: 115-130.</li>
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<li>Kriete G, Niehaus K, Perlick AM, Puhler A, Broer I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J 9: 809-818.</li>
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<li>Shukla P, Singh NK, Kumar D, Vijayan S, Ahmed I, et al. (2014) Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco. Funct Integr Genomics 14: 307-317.</li>
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<li>Goldman MH, Goldberg RB, Mariani C (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J 13: 2976-2984.</li>
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<li>Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, et al. (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27: 251-259.</li>
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<li>Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juarez P, Fernandez-del-Carmen A, et al. (2011) GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules. PLoS One 6: e21622.</li>
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Latest revision as of 03:27, 18 October 2014

Project > Modules > Biosafety


Biosafety


"With great powers comes great responsibility." – Benjamin Parker (Uncle Ben).



We know the importance of keeping genetically modified organisms under control. Ideally, modified genetic material should be unable to spread and organisms containing it should be easily distinguishable from wild type. Keeping this in mind, we created a biosafety module to be used in plants which will be available for future iGEM teams working with plants. Our biosafety module combines two biosafety strategies: identity preservation and male sterility.



Identity preservation enables an easy identification of the genetically modified organism. We expressed SlANT1 and SlJAF13 genes from tomato, which code for transcription factors that activate synthesis of anthocyanines [1], coloured compounds found in plants. The introduction of this element in the biosafety module generates differentially coloured plants (Figures 1 and 2). NRP-UEA-Norwich , and it is also used in their project “Green Canary”.



N benthamiana RoseaDark leaf

Figure 1.Anthocyanin accumulation in a N benthamiana plant.

Figure 2. Dark purple leaf with induced anthocyanin accumulation.


Male sterility makes impossible the dispersion of genetic material using pollen as the vehicle or by seeds result of self-pollination. In order to achieve this dispersion restriction, we integrated the active peptide of barnase, a RNAse from Bacillus amyloliquefaciens, (Biobricks accession code BBa_I716211) under the regulation of the tapetum-specific promoter TA29 [1]. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8]. We chose this strategy because it had been previously used in our laboratory with satisfactory results [9].


Our Biosafety module is our major present for future iGEM teams. It can be applied to a broad variety of new systems in future projects. We are certain that future iGEMers will realize how important biosafety is and they will incorporate our biosafety module in their systems.









References


  1. Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737-741.
  2. Cho HJ, Kim S, Kim M, Kim BD (2001) Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells 11: 326-333.
  3. Sa G, Mi M, He-Chun Y, Guo-Feng L (2002) Anther-specific expression of ipt gene in transgenic tobacco and its effect on plant development. Transgenic Res 11: 269-278.
  4. Shaya F, Gaiduk S, Keren I, Shevtsov S, Zemah H, et al. (2012) Expression of mitochondrial gene fragments within the tapetum induce male sterility by limiting the biogenesis of the respiratory machinery in transgenic tobacco. J Integr Plant Biol 54: 115-130.
  5. Kriete G, Niehaus K, Perlick AM, Puhler A, Broer I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J 9: 809-818.
  6. Shukla P, Singh NK, Kumar D, Vijayan S, Ahmed I, et al. (2014) Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco. Funct Integr Genomics 14: 307-317.
  7. Goldman MH, Goldberg RB, Mariani C (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J 13: 2976-2984.
  8. Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, et al. (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27: 251-259.
  9. Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juarez P, Fernandez-del-Carmen A, et al. (2011) GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules. PLoS One 6: e21622.


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