Team:Gothenburg/Safety

From 2014.igem.org

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<div class="imgRight" ><img style="height:200px" src="https://static.igem.org/mediawiki/2014/3/3b/Cs4Cas9firstcellcycle.png"/>
 
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<p>Figure 1. Dynamics of Csy4 and dCas9 expression during the first cell cycle</p>
 
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<h3>First cell cycle - the daughter cell resetter</h3>
 
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<p>Our system was designed in order to “reset” the age counter in newly born daughter cells. To accomplish this, one of the key components of the age counter, dCas9, is not produced in the very first cell cycle of daughter cells. As a consequence, the gRNA molecules that leak into the daughter cell during cell division won’t be able to induce the production of any fluorescent protein.
 
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As can be seen in fig. 1 we expect only Csy4 to be produced in the first cell cycle. To be noted, the G1-specific degradation tag inserted in both Csy4 and dCas9 is expected to trigger the respective degradation of the species at the end of the G1 phase. This feature can also be seen in fig1, in which Csy4 is rapidly depleted after the end of G1 phase.
 
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<h2>Safety in our iGEM project</h2>
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<p>Here you can find the Gothenburg team’s safety form, which was necessary to submit for the competition.</p>
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<h3>OurTraining</h3>
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<p><b>a) Have your team members received any safety training yet?</b></p>
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<p>Yes, we have already received safety training.</p>
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<p>Our daughter resetter not only impedes the production of fluorescent proteins induced by gRNA-dependent promoter but it also produces YFP, which becomes the characteristic color of the first cell cycle, together with the signal needed for the expression of CFP during the next cell cycle. As showed in fig. 2, during the first cycle YFP is constitutively produced by the endogenous daughter-cell specific promoter PDSE4. By design, together with the mRNA for YFP, also the 28-gRNA1-28 is produced. This is the signal the will last until the second cell cycle and, after being processed by CSY4 and bound to dCas9, will be able to induce the production of CFP. One non-ideality included in the model is the fact that CSY4 is present during the first cell cycle (fig. 1); as a consequence some 28-gRNA1-28 are turned into gRNA1 at the very moment of their creation. We had no way to test experimentally how long either form of gRNA lasts in the intracellular environment; this is the main source of uncertainty in our model. For the sake of simplicity, we chose a parametrization that would allow for both gRNA molelcules to survive degradation until the next cell cycle.
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<div class="imgRight" ><img style="height:340px" src="https://static.igem.org/mediawiki/2014/9/93/GRNA1-28firstcellcycle.png"/>
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<p>Figure 2. Dynamics of gRNA molecules during the first cell cycle.</p>
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<p><b>b) Please briefly describe the topics that you learned about (or will learn about) in your safety training.</b></p>
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<p>Work with dangerous compounds, Personal Safety Instructions (eye protection, clothes, gloves), Fire protection, Accident risks (cutting, splinters, hot equipment or liquid, burning materials, corrosive chemicals, inhalation of gases or vapors, electricity), Measures in case of accident.</p>
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<p><b>c) Please give a link to the laboratory safety training requirements of your institution (college, university, community lab, etc). Or, if you cannot give a link, briefly describe the requirements.</b></p>
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<p>Requirements before work in the lab: Read the Safety Instructions, Participating in an introductory lab tour including an introduction of the KLARA system, ordering and usage of autoclaves/centrifuges, Fill in the Risk Declaration form, Sign the Safety document and get a signature from the supervisor and from our lab manager.</p><br>
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<h3>Our local rules and regulations</h3>
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<p><b>a) Who is responsible for biological safety at your institution? (You might have an Institutional Biosafety Committee, an Office of Environmental Health and Safety, a single Biosafety Officer, or some other arrangement.) Have you discussed your project with them? Describe any concerns they raised, and any changes you made in your project based on your discussion.</b></p>
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<p>Marie Nordqvist. Yes. No concerns.</p>
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<p><b>b) What are the biosafety guidelines of your institution? Please give a link to these guidelines, or briefly describe them if you cannot give a link.</b></p>
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<a href"http://www.medarbetarportalen.gu.se/preventive_health_and_safety_work/arbetsmiljo-a-till-o/biosakerhet/"><p>Link</p></a>
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<p><b>c) In your country, what are the regulations that govern biosafety in research laboratories? Please give a link to these regulations, or briefly describe them if you cannot give a link.</b></p>
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<a href"http://www.folkhalsomyndigheten.se/amnesomraden/beredskap/biosakerhet-och-bioskydd/ "><p>Link</p></a><br>
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<h3>The Organisms and Parts that We Use</h3>
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<div style="padding-left:7.5%"><img  src="https://static.igem.org/mediawiki/2014/5/5e/Safetypngggg.png"/></div>
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<h3>Risks of Our Project Now</h3>
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<p><b>a)Risks to the safety and health of team members, or other people working in the lab:</b></p>
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<p>- PCR: upper temperatures can cause burns and should be dealt with carefully.
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<br>- Rotary shakers: glass shake flasks can break.
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<br>- Autoclave: high pressure build-up releases steam.
 +
<br>- Centrifuge: centrifugation is always hazardous since high speeds result in great forces. Great damage could result from improper handling of the centrifuge
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<br>- Danger of exposure to boiling water and high pressure steam in the autoclaving step.
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</p>
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<p><b>b) Risks to the safety and health of the general public (if any biological materials escaped from your lab):</b></p>
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<p>There are no risks with our constructs.</p>
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<p><b>c) Risks to the environment (from waste disposal, or from materials escaping from your lab):</b></p>
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<p>All biological waste is autoclaved and disposed by specialized companies</p>
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<p><b>d) Risks to security through malicious mis-use by individuals, groups, or countries:</b></p>
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<p>No.</p>
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<p><b>e) What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)</b></p>
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<p>- For PCR: do not open the thermal cycler when the machine is running.
 +
<br>- Rotary shakers: take care to clean up all the broken glass and be careful for sharp pieces of glass.
 +
<br>- Autoclave: use special autoclave gloves. Do not add chemicals to the water in the autoclave no sterilization of volatile or toxic substances. Do not seal containers, screw caps have to be loose , if glass explodes inside the autoclave - do not force the door to open. Protective gloves must insulate from heat exposure when handling items taken out of autoclave.
 +
<br>- Heat block: do not put water in the heat block. Do only use liquids in closed tubes.
 +
<br>- Centrifuge: NEVER run the centrifuge without proper training from responsible person. If a tube breaks in the centrifuge clean it up immediately! Dirt will eventually make the rotor corrode which may lead to serious accidents.
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<br>- Gel: Always operate under ventilation hood with gloves, avoid any contact with skin
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</p>
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<h3>Risks of our project in the future</h3>
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<p><b>a) What new risks might arise from your project's growth? (Consider the categories of risk listed in parts a-d of the previous question: lab workers, the general public, the environment, and malicious mis-uses.) Also, what risks might arise if the knowledge you generate or the methodsyou develop became widely available?</b></p>
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<p>As the projects aims to create an age counter, maybe the counter can be used to induce a toxin created only at a specific age, thus not detectable when the strain is of low age. We are sofar not creating any new methods.</p>
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<p><b>b) Does your project currently include any design features to reduce risks? Or, if you did all the future work to make your project grow into a popular product, would you plan to design any new features to minimize risks? (For example: auxotrophic chassis, physical containment, etc.) Such features are not required for an iGEM project, but many teams choose to explore them.</b></p>
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<p>No, there are no safety constructs included. No, we have not planned to include any.</p>
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Latest revision as of 20:09, 17 October 2014


TemplateUp

Biosafety

General safety in our lab

Firstly, in order to be allowed to work in the lab, everybody had to take part in a lab tour, when our lab manager introduced us into the safety and work rules. The following main parts were included in it:

  • Work with hazardous compounds

    • Hazardous substances are those that have injurious effect on health and environment. For example such substances can cause eye and skin irritation and damages or induce cancer. Hence in order to prevent ourself, our lab mates and the nature special handling and storage place is necessary. Taking care of the appropriate clothes and protection during the work with these compounds is also important.

  • Personal Safety Instructions

    • Lab coat, gloves and glasses were compulsory to wear during every experiment, what we have done. However special glasses are requirements when UV light and blue light are used. Such glasses are provided in our lab, near the certain instruments.

  • Accident risks during the work

    • The main accident risks can be mechanical injuries, burns and corrosion, poisoning, and electric shocks.
    • Cutting: Never use damaged glasswear! Glass should be collected in designated boxes to avoid injury to the cleaning personnel.
    • Burns and corrosion: Bunsen burners have almost invisible and silent flame so it is a hazard. The most common accidents in a laboratory is fire causing flammable solvents. Since our department work with large amounts of such liquids it is important that all person is aware of how to handle such a fire.
    • Poisoning: It can occur by inhalation of gases or fumes, or through the skin.
    • Electric shocks: If a person has gotten in contact with an AC line there is a severe muscle convulsion that makes the person unable to get loose from the line. Such cases are extremely dangerous.

  • Fire protection

    • Equipment for fire extinction is found in all corridors. This equipment consists of water hoses, portable fire extinguishers (CO2 or powder), and fire cloths. There are also heat detectors and smoke detectors in the building in case of a fire.



    Except from the Introductory Lab Tour, all of the team members were obliged to:
    • Read the Safety Instructions of the lab
    • Fill in a Risk Declaration form
    • Sign the Safety document and get a signature from the supervisor and from our lab manager

Video

A video was established by our lab manager (Marie Nordqvist) about the main risks in our laboratory


Safety in our iGEM project

Here you can find the Gothenburg team’s safety form, which was necessary to submit for the competition.

OurTraining

a) Have your team members received any safety training yet?

Yes, we have already received safety training.

b) Please briefly describe the topics that you learned about (or will learn about) in your safety training.

Work with dangerous compounds, Personal Safety Instructions (eye protection, clothes, gloves), Fire protection, Accident risks (cutting, splinters, hot equipment or liquid, burning materials, corrosive chemicals, inhalation of gases or vapors, electricity), Measures in case of accident.

c) Please give a link to the laboratory safety training requirements of your institution (college, university, community lab, etc). Or, if you cannot give a link, briefly describe the requirements.

Requirements before work in the lab: Read the Safety Instructions, Participating in an introductory lab tour including an introduction of the KLARA system, ordering and usage of autoclaves/centrifuges, Fill in the Risk Declaration form, Sign the Safety document and get a signature from the supervisor and from our lab manager.


Our local rules and regulations

a) Who is responsible for biological safety at your institution? (You might have an Institutional Biosafety Committee, an Office of Environmental Health and Safety, a single Biosafety Officer, or some other arrangement.) Have you discussed your project with them? Describe any concerns they raised, and any changes you made in your project based on your discussion.

Marie Nordqvist. Yes. No concerns.

b) What are the biosafety guidelines of your institution? Please give a link to these guidelines, or briefly describe them if you cannot give a link.

Link

c) In your country, what are the regulations that govern biosafety in research laboratories? Please give a link to these regulations, or briefly describe them if you cannot give a link.

Link


The Organisms and Parts that We Use

Risks of Our Project Now

a)Risks to the safety and health of team members, or other people working in the lab:

- PCR: upper temperatures can cause burns and should be dealt with carefully.
- Rotary shakers: glass shake flasks can break.
- Autoclave: high pressure build-up releases steam.
- Centrifuge: centrifugation is always hazardous since high speeds result in great forces. Great damage could result from improper handling of the centrifuge
- Danger of exposure to boiling water and high pressure steam in the autoclaving step.

b) Risks to the safety and health of the general public (if any biological materials escaped from your lab):

There are no risks with our constructs.

c) Risks to the environment (from waste disposal, or from materials escaping from your lab):

All biological waste is autoclaved and disposed by specialized companies

d) Risks to security through malicious mis-use by individuals, groups, or countries:

No.

e) What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)

- For PCR: do not open the thermal cycler when the machine is running.
- Rotary shakers: take care to clean up all the broken glass and be careful for sharp pieces of glass.
- Autoclave: use special autoclave gloves. Do not add chemicals to the water in the autoclave no sterilization of volatile or toxic substances. Do not seal containers, screw caps have to be loose , if glass explodes inside the autoclave - do not force the door to open. Protective gloves must insulate from heat exposure when handling items taken out of autoclave.
- Heat block: do not put water in the heat block. Do only use liquids in closed tubes.
- Centrifuge: NEVER run the centrifuge without proper training from responsible person. If a tube breaks in the centrifuge clean it up immediately! Dirt will eventually make the rotor corrode which may lead to serious accidents.
- Gel: Always operate under ventilation hood with gloves, avoid any contact with skin

Risks of our project in the future

a) What new risks might arise from your project's growth? (Consider the categories of risk listed in parts a-d of the previous question: lab workers, the general public, the environment, and malicious mis-uses.) Also, what risks might arise if the knowledge you generate or the methodsyou develop became widely available?

As the projects aims to create an age counter, maybe the counter can be used to induce a toxin created only at a specific age, thus not detectable when the strain is of low age. We are sofar not creating any new methods.

b) Does your project currently include any design features to reduce risks? Or, if you did all the future work to make your project grow into a popular product, would you plan to design any new features to minimize risks? (For example: auxotrophic chassis, physical containment, etc.) Such features are not required for an iGEM project, but many teams choose to explore them.

No, there are no safety constructs included. No, we have not planned to include any.

Retrieved from "http://2014.igem.org/Team:Gothenburg/Safety"