Microinjecting eggs with antisense Morpholino oligonucleotides (MO) to knock down specific genes of interest. MO injection is the standard technique for eliminating gene expression in zebrafish embryos. This technology provides a way of rapidly determining gene function in a vertebrate organism and can be accomplished in a matter of weeks.
Microinjecting eggs with RNA in order to over-express wild-type, mutant and chimeric genes. RNA injection is the standard technique for ubiquitously expressing a gene of interest. The level of over-expression (dose) can be controlled by adjusting the amount of injected RNA.
Microinjecting eggs with DNA to express wild-type, mutant and chimeric genes in a mosaic pattern in the embryo. Transient expression from injected DNA is highly mosaic in zebrafish. This is useful if the introduced DNA product is likely to be toxic. The mosaic expression can be modulated by the choice of vector, and by adjusting the amount of injected DNA. Using this technique, the effect of injected DNA on cellular behaviors can be studied in the context of unaltered cells in the developing embryo.
Screening for toxicity/drugs using 96-well format. Zebrafish embryos can be screened for alterations in phenotype or toxicity using environmental toxins or small molecule libraries. In addition, transgenic reporter lines can be screened for effects on specific molecules or organ systems.
Analyzing phenotypes (including morphology, gene expression, cell cycle, histochemistry, etc.) is facilitated by the ZSR Director who has over fifteen years of experience working with zebrafish embryos.
Generation of transgenic zebrafish by microinjecting eggs with DNA constructs. The injected embryos are raised to adulthood and mated. Transmission of the transgene is determined (usually by the presence of a fluorescent marker) in the progeny of the founding fish. Embryos that carry the transgene are raised to adulthood and intercrossed to establish the transgenic line.
Transplantating cells is used to determine whether a gene product functions in the cell autonomously, or if it influences, or is influenced by, its cellular environment.
Transplantating human tumor cells into embryonic or adult zebrafish makes use of the transparency of zebrafish to visualize the behavior of the tumor cells in the live animal. In addition, the environment of the embryo can be manipulated by using mutant or transgenic fish, or by exposure to drugs or toxins simply added to their water.
Designing custom experimental protocols to fit specific requirements for the research needs of individual investigators.
Training is provided at all levels and on all aspects of zebrafish research.
Consulting on all aspects of zebrafish research is provided.