SAFE PROJECT DESIGN

    Although centromeric retrotransposon (CR) elements are not viral, we decided not to pursue cloning the entire sequence. While ~50% of the human genome is composed of transposable elements (International Human Genome Sequencing Consortium, 2001), cloning the entire sequence of a centromeric retrotransposon of Zea mays may pose various risks. The potential adverse effects of retrotransposons in organisms have not been demonstrated as of yet. However, retrotransposable elements are still ‘parasitic’ in that they can reintegrate themselves into the genome and persist for long periods of time. Thus, we took necessary precautions and used only the partial sequence, omitting the integrase portion, to limit possible risks of retrotransposon exposure.

    Initially, we thought of using plants in our experiment to observe virus-like particle assembly for DNA transport with fluorescent tags. However, exposure to portions of these retrotransposon elements and the packaged contents of a virus-like particle may, in an extreme case, lead to uncontrolled dispersion of genetically modified DNA. With much of the mechanistic details of its reinsertion unknown, we thus decided to work with our elements in vitro. Non-pathogenic E. coli strains (BL21 and DH5a) were used to clone our partial CR construct and all members were trained to work safely in the lab.

PREPARING FOR SAFE LAB WORK

    All members of the lab received thorough safety training that covers content of the UH Manoa Chemical Hygiene Plan, physical and health hazards of chemicals, and methods to detect hazardous conditions. Members were trained by UH Manoa’s Environmental Health & Safety Office (EHSO) education coordinator Leimomi Kekina and biosafety and biological waste management professional Stephen Case.

SAFETY IN THE LAB

    Members of the lab made sure to always wear proper personal protective equipment (PPE) and work in the proper environment when dealing with potentially hazardous chemicals. Toxic and carcinogenic reagents were limited at every time possible. For example, instead of using ethidium bromide to stain our agarose gels, we used gel red, a much safer alternative. We brought potentially toxic reagents to the hood and efficiently worked with them, trying to limit our exposure to the chemicals.

REFERENCES

International Human Genome Sequencing Consortium. (2001). Initial sequencing and analysis of the human genome. Nature, 409(6822), 860–921. https://doi.org/10.1038/35057062