Difference between revisions of "Team:EPF Lausanne/Safety"

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<h2>Safety in iGEM</h2>
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<p>Please visit <a href="https://2015.igem.org/Safety">the main Safety page</a> to find this year's safety requirements & deadlines, and to learn about safe & responsible research in iGEM.</p>
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<p>On this page of your wiki, you should write about how you are addressing any safety issues in your project. The wiki is a place where you can <strong>go beyond the questions on the safety forms</strong>, and write about whatever safety topics are most interesting in your project. (You do not need to copy your safety forms onto this wiki page.)</p>
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<h4>Safe Project Design</h4>
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<p>Does your project include any safety features? Have you made certain decisions about the design to reduce risks? Write about them here! For example:</p>
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<li>Choosing a non-pathogenic chassis</li>
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<li>Choosing parts that will not harm humans / animals / plants</li>
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<li>Substituting safer materials for dangerous materials in a proof-of-concept experiment</li>
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<li>Including an "induced lethality" or "kill-switch" device</li>
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<h4>Safe Lab Work</h4>
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<p>What safety procedures do you use every day in the lab? Did you perform any unusual experiments, or face any unusual safety issues? Write about them here!</p>
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<h4>Safe Shipment</h4>
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<p>Did you face any safety problems in sending your DNA parts to the Registry? How did you solve those problems?</p>
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        <h2>Safety</h2>
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      <h1>General safety measures</h1>
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      <p>As every person working with living microorganisms we had to follow a set of simple rules. First of all always wear a labcoat and gloves at any given time in the laboratory. Before starting and after finishing to work on a bench decontaminate the place with ethanol. All our waste and biowaste were treated accordingly to the legislation in our country.</p>
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      <h1>Special safety needs for our project</h1>
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      <p>Our project is based on the dCas9, a derived form of the very polemical CRISPR-Cas9 protein which raised safety issues concerning the “off target effects” (when the Cas9 is guided to and cuts an unwanted sequence in the genome). These effects led us to think about some ways to minimize the risks of unwanted targeting, both in the genome of E.coli and S. cerevisiae we are working with. Even though we won’t introduce Cas9 or dCas9 into a human cell we wanted to check if there was a risk of gene activation/inhibition if we got a large quantity of these proteins into our body. In our case (with dCas9), the risk is the following: if a guide RNA causes dCas9 to bind a regulatory region of gene in an unwanted fashion, it might be deadly for the organism.</p>
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      <h1>What did we do to reduce the risk?</h1>
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      <p>To assess these issues we designed <a href="https://2015.igem.org/Team:EPF_Lausanne/Software">a program</a> that automatically blasts the guide RNA (gRNA) plus de PAM sequence (either CCn at the beginning or nGG at the end) against the genome of E. coli, S. cerevisiae and the human genome. When we had the result the program would give us the sequences that are not in the organisms. Note that we wanted to work with completely synthetic gRNA to have as little interference as possible. Future iGEM teams could use our software to read their BLAST results and to draw conclusions of the results that could maybe result in less “off-target effects”.</p>
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Latest revision as of 09:07, 18 September 2015

EPFL 2015 iGEM bioLogic Logic Orthogonal gRNA Implemented Circuits EPFL 2015 iGEM bioLogic Logic Orthogonal gRNA Implemented Circuits

Safety

General safety measures

As every person working with living microorganisms we had to follow a set of simple rules. First of all always wear a labcoat and gloves at any given time in the laboratory. Before starting and after finishing to work on a bench decontaminate the place with ethanol. All our waste and biowaste were treated accordingly to the legislation in our country.

Special safety needs for our project

Our project is based on the dCas9, a derived form of the very polemical CRISPR-Cas9 protein which raised safety issues concerning the “off target effects” (when the Cas9 is guided to and cuts an unwanted sequence in the genome). These effects led us to think about some ways to minimize the risks of unwanted targeting, both in the genome of E.coli and S. cerevisiae we are working with. Even though we won’t introduce Cas9 or dCas9 into a human cell we wanted to check if there was a risk of gene activation/inhibition if we got a large quantity of these proteins into our body. In our case (with dCas9), the risk is the following: if a guide RNA causes dCas9 to bind a regulatory region of gene in an unwanted fashion, it might be deadly for the organism.

What did we do to reduce the risk?

To assess these issues we designed a program that automatically blasts the guide RNA (gRNA) plus de PAM sequence (either CCn at the beginning or nGG at the end) against the genome of E. coli, S. cerevisiae and the human genome. When we had the result the program would give us the sequences that are not in the organisms. Note that we wanted to work with completely synthetic gRNA to have as little interference as possible. Future iGEM teams could use our software to read their BLAST results and to draw conclusions of the results that could maybe result in less “off-target effects”.

EPFL 2015 iGEM bioLogic Logic Orthogonal gRNA Implemented Circuits