Week 4: Reading Continued

Welcome back to my blog! 

 

This week I continued to read up on transgenic plants. 

After reading the paper, “Public health issues related with the consumption of food obtained from genetically modified organisms” by Paparini & Romano-Spica, I wanted to learn more about gene transfer and what health and environmental risks are associated with genetically engineered crops. I wanted to learn more about this topic because one argument against genetically modified organisms is that eating GMOs could change one’s genome and that the ecosystem could be disrupted by the introduction of modified plants. From my reading, I deduced that it is highly unlikely for transgenes from GMO food to change the genome of humans [1]. However, long-term environmental and human health risks are hard to assess due to the novelty of different GMOs and the rapid change in the industry. As such, GMOs should be regularly monitored. GMOs can impact the state of soil microbiomes, insect populations, and more [2, 3, 4]. I am curious to see how the impact of GMOs on the environment is different than non-GMO and organic farming.

In the lab:

Last week, the DNA extraction failed, so this week we tried again! On Tuesday, Dr. Lindsay and I went to the farm to collect leaf samples from our CRISPR mutants. We then went back to the lab and began the DNA extraction procedure. 

 

The first step was to crush down the sample, so we froze our samples using liquid nitrogen and then used a TissueLyser machine to crush our samples into a fine powder. The next steps of the procedure were to separate the plant’s DNA from the rest of the plant material. Lastly, we set up a PCR reaction using the extracted DNA as the template and let it run overnight.

The next day we ran the PCR product on a gel, and we were glad to see that the samples looked good! The DNA extraction worked! We decided to double-check a few of the DNA samples we planned to send into sequencing. After performing the PCR, I sterilized the PCR product, and then because I had to leave, my mentor used the nanodrop machine to quantify the sterilized PCR product concentration of the samples and ran them on a gel.

 

Also since the LR reaction for last week was successful, my mentor and I put our transformed e.coli into agrobacteria via electroporation, then plated the agrobacteria. On Friday after a lab meeting and watching a Ph.D. defense presentation, my mentor and I performed a colony PCR on our transformed agrobacteria from this week and then ran the samples on a gel. The gel showed us that the transformation was successful!

 

Sources:

1. Paparini, A., & Romano-Spica, V. (2004). Public health issues related with the consumption of food obtained from genetically modified organisms. Biotechnology annual review, 10, 85–122. https://doi.org/10.1016/S1387-2656(04)10004-5
2. Kuvshinov, V., V, Koivu, K., Kanerva, A., & Pehu, E. (2001). Molecular control of transgene escape from genetically modified plants. Plant science: an international journal of experimental plant biology, 160(3), 517–522. https://doi.org/10.1016/s0168-9452(00)00414-3
3. EFSA Panel on Genetically Modified Organisms (GMO), Naegeli, H., Bresson, J. L., Dalmay, T., Dewhurst, I. C., Epstein, M. M., Firbank, L. G., Guerche, P., Hejatko, J., Moreno, F. J., Nogue, F., Rostoks, N., Sanchez Serrano, J. J., Savoini, G., Veromann, E., Veronesi, F., Casacuberta, J., De Schrijver, A., Messean, A., Patron, N., … Mullins, E. (2021). Evaluation of existing guidelines for their adequacy for the molecular characterisation and environmental risk assessment of genetically modified plants obtained through synthetic biology. EFSA journal. European Food Safety Authority, 19(2), e06301. https://doi.org/10.2903/j.efsa.2021.6301
4. Pepoyan, A. Z., & Chikindas, M. L. (2020). Plant-associated and soil microbiota composition as a novel criterion for the environmental risk assessment of genetically modified plants. GM crops & food, 11(1), 47–53. https://doi.org/10.1080/21645698.2019.1703447