Progress 10/01/15 to 09/30/18
Outputs
Target Audience:Food industry including food-packaging industry and consumers of those products are target audience and will benefit from this study. It will provide food-packaging industry and chemical industry to make a decision on selecting safer chemical alternatives and type of food packaging to use intended for certain application. If NYS industry adopts the screening tools established by this study, it would place NYS food industry ahead of the game and boost consumer confidence in products made in NYS. The screening tool can also be applied to food packaging and food products imported from foreign countries where regulations may not be as rigorous as in the U.S. Consumers will be provided with more information, which type of food packaging or containers are safe or unsafe (if any), especially when containing food intended for babies, small children, and pregnant women. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project has provided training and professional development opportunities for one undergraduate student, graduate student, two technical staff (one is a recent Cornell BS graduate), and a postdoc. How have the results been disseminated to communities of interest?Two poster presentations were given at international society meetings and the abstracts of these meetings are made available to the public. Two publications are currently under preparation. An undergraduate student have presented a poster titled "Generation plastic: Is BPA determining adulthood?" at the Cornell President Inauguration event. This was a campus wide event highlighting the scholarship across campus. The PI has also presented the importance of ensuring safety of food packaging to food companies that occasionally visits the Department and possible use of zebrafish as high-throughput screening model. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
During the first 2 years of the project, we successfully generated four transgenic reporter lines with genes tagged with fluorescent proteins using CRISPR/CAS9 technology. This involved systematic steps in developing targeting DNA, embryo injections and screening to identify successful genetic targeting. In the final reporting period, we reached several additional milestones. First, we made one additional transgenic line of zebrafish using Tol2 technique for a genetic locus that proved difficult to target using CRISPR/CAS9 technology. Thus, over the period of this project, a total of 5 transgenic lines were generated and colonies established in our lab. Second, these lines were used to assess toxicity of food packaging migrants as complex mixture as well as for specific compounds of interest, such as BPA alternatives. We tested the sensitivity of one of these lines designed to detect estrogenic compounds. Exposure to different concentration of 17β-estradiol (E2; 0 - 625 nM) revealed induction of hepatic EGFP expression in larvae exposed to 2.5 nM E2 as early as 3-days post-fertilization. Induction of hepatic EGFP expression was observed as low as 10 uM with bisphenolic compounds, which were in line with reported estrogenic potential of bisphenol compared to endogenous estrogens. In comparison to two previously published reports attempting to generate reporter lines similar to ours (without CRISPR/Cas9 technology, our transgenic line was of several magnitudes of order more sensitive. This represents a significant advancement towards application for this product. . Finally, we have also experimented methods to make the toxicology screening semi-automatic for use in high-throughput settings, to test toxicity of leachate from food packaging material as well as any other potentially toxic compounds. This is the first study to use the novel CRISPR/CAS9 technology to develop fluorescently labeled zebrafish to detect endocrine disrupting compounds. We have presented the findings at an international toxicology conference and our results have been well-received with interest from other scientists in academia and industry, trainees, and regulatory agencies. A manuscript is currently under preparation to publish these results in a peer-reviewed journal to broadly disseminate this research. The developed transgenic zebrafish lines would also find applications as a unique tool for other toxicology studies, not just for food packaging migrants - thus having a much broader impact on protecting safety of food, water, products, and the environment.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Abdel-moneim, A., and Mukai, M. (2019) In vivo hight-throughput screening system for estrogenic endocrine disruptors using a fluorescently-labeled zebrafish line , The Toxicologist: Supplement to Toxicological Sciences, 168 (1), no. 3151.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Abdel-moneim, A., and Mukai, M. (2018) Development of an in vivo fluorescent reporter assay for high-throughput screening of estrogenic compounds using CRISPR/Cas9 system in zebrafish , The Toxicologist: Supplement to Toxicological Sciences, 162, no. 3547.
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience:Food industry including food-packaging industry and consumers of those products are target audience and will benefit from this study. It will provide food-packaging industry and chemical industry to make a decision on selecting safer chemical alternatives and type of food packaging to use intended for certain application. If NYS industry adopts the screening tools established by this study, it would place NYS food industry ahead of the game and boost consumer confidence in products made in NYS. The screening tool can also be applied to food packaging and food products imported from foreign countries where regulations may not be as rigorous as in the U.S. Consumers will be provided with more information, which type of food packaging or containers are safe or unsafe (if any), especially when containing food intended for babies, small children, and pregnant women. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project has provided training and professional development opportunities for one undergraduate student, graduate student, two technical staff (one is a recent Cornell BS graduate), and a postdoc. How have the results been disseminated to communities of interest?An undergraduate student have presented a poster titled "Generation plastic: Is BPA determining adulthood?" at the Cornell President Inauguration event. This was a campus wide event highlighting the scholarship across campus. The PI has also presented the importance of ensuring safety of food packaging to food companies that occasionally visits the Department and possible use of zebrafish as high-throughput screening model. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we plan to complete the generation of additional lines and continue on with the experiments as proposed in the original proposal.
Impacts
What was accomplished under these goals?
We have now made total of 4 transgenic lines of zebrafish with genes tagged with fluorescent proteins which allows the monitoring of toxicity of food packaging migrants, such as bisphenol A and its alternatives. These transgenic fish provide an excellent tool for screening toxic compounds where toxicological effects can be monitored in a live animal at multiple concentrations and in different developmental stages. We have tested these lines with estrogenic compounds to test the sensitivity of the line as a indicator screen. Generation of additional lines are also underway.
Publications
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Progress 10/01/15 to 09/30/16
Outputs
Target Audience:All industry involved in the manufacturing and sales of food products and consumers are target audience and will benefit from this study. It will also provide the food-packaging industry with information on which BPA alternatives are safe to use. If NYS industry adopts the screening tools established by this study, it would place NYS food industry ahead of the game and boost consumer confidence in products made in NYS. The screening tool can also be applied to food packaging and food products imported from foreign countries where regulations may not be as rigorous as in the U.S. Consumers will know which type of food packaging is safe or unsafe (if any), especially for developing children and pregnant women. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project has provided training and professional development opportunities for one graduate student and one undergraduate student. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?For the next reporting period, we plan to make 1-2 additional transgenic lines and continue on with the experiments as proposed in the original proposal.
Impacts
What was accomplished under these goals?
We have made 2 trangenic lines of zebrafish with genes tagged with green fluorescent protein which allows the monitoring of toxicity of food packing migrant such as bisphenol A. These transgenic fish provide an excellent tool for screening toxic compounds where toxicological effects can be monitored in a live animal at multiple concentrations and in different developmental stages. We are also now in the process of testing the toxicity of some BPA alternatives to see if they have similar effect as BPA or not.
Publications
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