What safety practices we should have in place prior to transforming or transfecting plants?
At the BioCurious hackerspace in the SF bay area, we have a group who would like to transform/transfect plants. We currently have only transformed bacteria and yeast before. The group is looking at two approaches, using Agrobacterium and using a gene gun. We're looking at what safety practices we should have in place prior to doing this. Some specific questions:
Are tight controls preventing transformed plant material not leaving the lab before being autoclaved necessary? What risk would be present?
How hazardous is a gene gun, especially if particles containing intended plant vectors were accidentally shot at someone's hand/body?
Are there other safety considerations we should be aware of before proceeding with this work?
Answer from a Biosafety Officer:
February 3, 2013
1. Are tight controls preventing transformedplant material not leaving the lab before being autoclaved necessary? What riskwould be present?
In the case of transformed or transfectedplants the recommendations for inactivation of plant materials prior to leaving the laboratory are in place to protect the environment by preventing transgenic plant materials from entering the food chain, and/or unintentionally transferring the transgene under study to plants in the environment.
Any Agrobacterium cultures should be treated with 1 part Clorox bleach to 9 parts of culture for a minimum of 20 minutesprior to disposal.
In most research facilities an autoclave is available to inactivate plant materials prior to removal from the lab. If an autoclave is not available, plant material may be inactivated by heating to 85-100 degrees C in a box made of heat-resistant materials using electric heating coils (known as soil sterilizers in the greenhouse industry).
If plants without seeds are used in the experiment they may be inactivated by withholding water until the plant is fully desiccated prior to disposal. If plants with seeds are used in the experiment, then seeds should be collectedfor inactivation prior to disposal as well.
2. How hazardous is a gene gun, especially if particles containing intended plant vectors were accidentally shot at someone'shand/body?
Gene guns have been routinely used to deliver vectors to the skin in research studies, and particles accelerated from a genegun can penetrate multiple layers of tissue. Without specific information on the transgene and plant vector that will be delivered, it is difficult to say exactly what might happen if plant vectors were introduced into human skin, but the best practice would be to avoid exposure.
Do not point the gene gun at people or parts of your body. Plant specimens should be secured using clamps and/or stands during the gene delivery process. Users should not hold specimens to be transformed while the gene gun is being used.
Gene guns produce brief, high intensity sounds during use, and repeated exposure could lead to hearing loss. Hearing protection should be worn by any personnel in the vicinity of the gene gun while it is being used.
Gene guns also require the use of compressed gasses. Be sure to secure all compressed gas cylinders by mounting a bracket or strap to a sturdy bench or using a wall bracket to prevent the cylinder from falling over.
3. Are there other safety considerations weshould be aware of before proceeding with this work?
Seeds from transgenic plants can be collected by placing fine mesh bags around the flower heads prior to disposal. Plant pots can be kept on a tray lined with absorbent materials to catch stray seeds and pots, trays, and other solid materials can be soaked in a 1:10 dilution of bleach after use.