Overview

Safety is a big concern in all laboratory work, which is why we would like to share some of the precautions we took in order to minimize risks to the environment and ourselves. When working with genetically modified organisms, we must ensure that they do not escape into the environment. For this reason our Chlamys cannot survive in the wild.

Before we start

Before working in the lab, each lab member participated in a lab safety briefing conducted by qualified supervisors. The briefing included general lab safety and instructions on how to handle dangerous situations. This briefing is mandatory for anyone who wants to work in the laboratory and official documents must be signed proving that the briefing was conducted.

Our Laboratory

We work in S1 (German biosafety level 1) laboratories, this biosafety level is appropriate for work with organisms that do not pose any environmental or human risks. The following attire is required in these labs:

Lab coats
Long pants, covering knees and shins
Closed shoes, toes must be covered
Long hair must be tied up
When working with dangerous chemicals, such as boiling agar or acrylamid, it is mandatory to wear safety goggles and gloves

This is a picture of an exemplary student wearing glasses, gloves and his labcoat. He is also wearing long pants and closed shoes with his hair tied up.

In general we recomend good skin coverage to avoid injury, clothes can be washed and replaced our skin is precious. Common rules include prohibiting pets and children from entering. Moreover, eating, drinking, chewing gum or applying makeup is also forbidden. In order to avoid contaminations of all kinds, electronic devices are not to be placed on the bench, this includes smartphones, tablets and laptops. This reduces the risk of contaminating our samples with outside bacteria but also hinders taking contaminants out of the lab and into our homes or our environment. Additionally, we wash and disinfect our hands before entering and leaving the lab. While working in a lab it is critical to use common sense, especially in the case of an emergency, our laboratory is well fitted with eye wash station, shower stations, fire alarms, fire extinguisher and fire blankets among other safety features, students who wish to work in the lab must be made familiar with these features in order to feel confident and calm during an emergency. In case of a fire, there is a meeting point outside of the building where we must all meet in order to perform a head count and ensure everyone' safety.

Sterile Bench

Working with E. coli or Chlamydomonas reinhardtii involves working under a sterile hood, often in the radius of an open flame. For this reason it is imperative that we practice good habits and be extra careful of safety regulations while operating around a burning flame. Besides the general rules of keeping the hood sterile and avoiding contamination there are additional safety rules that must be followed while working under the hood:

An open flame must never be left unattended
Ethanol must not be used in the form of aerosol near the flame ie with a spray bottle
The airflow must not be blocked by contaminants or aerosol as this will end up in the air and in our lungs
All waste that comes in bacteria or algea must be sorted into auoclave trash

Working under the hood is a special kind of lab work and it requires practice, clear and particular movements and good habits.

Chlamydomonas reinhardtii

Our adorable unicellular green algae, Chlamydomonas reinhardtii or Chlamy, is in a risk group 1, white listed, this means that it is non-toxic for humans and for the environment. It is also rated as GRAS which means Generally Recognized As Safe by FDA standards. Our lab has a lot of experience with this organism. Furthermore it is very well studied, so we are confident we can safely handle this organism in our labs. We specifically use a Chlamy stain with weakened cell walls which means it can only weakly survive even in optimal conditions.

Our parts

CYPs are also safe and our created constructs have no selection advantages, so if our algae ever got out with these CYP inserts, they cannot harm wild algae. The parts have very Chlammy specific codon usage and all constructs have introns that are Chlammy specific, which means they are not correctly spliced by other species and would lead to nonsense proteins.

Experiments

These rules must be actively followed in our day to day work in the lab, however, some experiments remain inherently dangerous. We wanted to avoid using dangerous subtances and methods as much as possible, sometimes small differences can make the biggest impact. One example, is our circumvention of using ethidium bromide, a known DNA intercalating agent and carcinogen. We opted instead for HDGreen, a safer alternative. The safety of HDGreen is tested and outlined by the manufacturer Intas in their report. Another example, is our avoidance of using carbon monoxide as an activity assay for CYPs. Carbon monoxide or CO is a tasteless, odorless and colorless gas, which means it can be inhaled unconsciously, it is so toxic it can lead to tachycardia and tachypnea, after prolonged consumption it can even be lethal.¹ Instead, we have developped our own methode using Native PAGE which can be seen in our Contributions.

Hanley ME, Patel PH. Carbon Monoxide Toxicity. [Updated 2023 Jan 23]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK430740/