Influence of the microinjection process on the survival of embryos Zebrafish (Danio rerio) in the production of transgenic fish

Fish belong to convenient objects of research, tk. they are not expensive in the content, they have transparent shells that allow easy analysis of developmental stages, produce a lot of fertilized eggs required for introduction of genes. The purpose of this work was to study the effect of the introduction of genetic constructs by microinjection on the viability of transgenic model embryos and larvae of fish with fluorescent reporter genes. Using a micromanipulator, two different gene constructs containing the reporter gene GFP were introduced, as a control, distilled water was microinjected. The advantage of these constructs is the intravital expression of the green fluorescent protein under UV irradiation and FITC filter application. Analysis of developmental stages, morphological changes in fish, and gene expression were studied on a Lumar 12 stereo microscope by Zeiss. Within a day, analysis of the early stages of fish development both in control and in experimental groups showed the presence of normal and abnormal forms of development. In addition, non-viable organisms have been identified in all embryo groups studied. Thus, it is the first day that is the main critical period of embryo survival. A total of 335 embryos were analyzed, among them in control 60, after the introduction of water 111, after the introduction of the pCX-EGFP construct 126 and 38 after the pCEEGPF. The different viability of embryos and larvae in control and using different genetic constructs is shown for the first time. The critical periods of life of the early stages of development in the experimental groups of fish were first revealed. Transgenic embryos and Danio rerio larvae were obtained with the mosaic nature of the expression of the green fluorescent protein (GFP). It should be noted that the influence of gene modifications on the homeostasis and the life span of genetically modified organisms at the population, cellular, and molecular levels has not been sufficiently studied. Further research is needed on the development and life span of transgenic organisms.

transgenic animals, plasmids, reporter genes, gene expression, microinjection, genetic and construction, embryonic-larval development, zebrafish, survival

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