a Part of Collaborative Innovation Center of Genetics and Development
Dr. Yan Pi | 皮妍
Zebrafish represent an attractive vertebrate model in which phenotypes at the molecular, cellular and behavioral levels can be rapidly analyzed. The species is demonstrably similar to other mammalian models in functional studies and toxicity testing. Our lab focuses on using zebrafish together with CRISPR technology, a new genome-editing tool, as a platform for highly efficient analysis of genes related to neuropsychiatric disorders. We employ CRISPR technology to efficiently generate zebrafish mutations in selected gene families to illustrate the function of these genes in brain development and social behavior. These models also serve as invaluable tools for future drug screening to identify potential therapeutic candidates.
The genetics module I designed for BIOS will first introduce the developmental process of zebrafish. The application of zebrafish in environmental, drug, and food safety and toxicity assessment will be discussed and such experiments will be designed and performed according to the interest of each student. Students will have the opportunity to observe firsthand aberrant development and design basic experiments of their own.
Zebrafish pigment mutants. A wild-type zebrafish (top) is shown for comparison. The mutants lack black pigment in their melanocytes because they are unable to synthesize melanin properly. It is easy to know that the pigmentation gene has been mutated by the CRISPR/Cas9 system. (Proc Natl Acad Sci U S A. 2013; 110(34): 13904–13909)
Zebrafish embryonic development is very rapid, with precursors to all major organs appearing within 36 hours of fertilization. The embryos are relatively large, robust, and transparent, allowing us to see development outside the mother. After a few months, the adult fish reaches reproductive maturity.
In situ hybridization – cmlc2 gene in adult zebrafish. In situ hybridization is used to reveal the location of specific nucleic acid sequences on chromosomes or in tissues, a crucial step for understanding the organization, regulation, and function of genes.
Analysis of zebrafish embryos in multi-well plates. Zebrafish has long served as a model organism for high-throughput drug screening. Because its embryo is small in size, optically transparent, and can be kept alive in multi-well plates for several days without the need for additional nutrients, it is also a very easy organism to work with. In addition, zebrafish are easy to raise, yield large numbers of offspring, and the instrumentation required to perform screens has been reported to be less extensive than that found in many other animals. Methodology for automated live imaging and analysis of compound-treated zebrafish embryos using multi-well imaging in combination with artificial intelligence-based analysis is capable of measuring changes in fine structure not visible to the human eye. (Dev Dyn. 2009; 238(3): 656–663)
