Abstract
Genetically modified (GM) animals are necessary to solve the global problems of humanity related to nutrition and health. Rabbits, as laboratory, domestic and farm animals, occupy a special niche in research. GM rabbits are promising as bioreactors for producing biologically active (BA) proteins with milk or blood, and are in demand in Biomedicine as biomodels of diseases. To date, many GM rabbits-biomodels, producers of recombinant proteins have been created in the world using CRISPR/Cas9 technology. All-Russian Research Institute of Animal Physiology, Biochemistry and Nutrition has experience in obtaining transgenic rabbitsproducers of human BA proteins with milk by microinjecting recombinant DNA into zygote pronuclei. The possibility of site-specific modification of the rabbit whey acidic protein (WAP) gene using CRISPR/Cas9 technology is discussed. A DNA matrix containing homology arms to the WAP rabbit gene and site-specific CRISPR/Cas9 components in plasmid form were obtained. Microinjections of rabbit zygotes were performed and embryo survival was evaluated in vitro. The efficiency of using the green fluorescent protein gene under the cytomegalovirus promoter in the DNA matrix as an indicator of homologically directed repair was evaluated. This work can be useful for obtaining rabbits that produce with milk BA protein instead of WAP.
Highlights
The main goals of obtaining genetically modified animals are to change the characteristics of animals, to acquire new opportunities for them, to eliminate some negative qualities in the desired direction for humans
It has been used for studying human diseases for a long time, and it is used as a biomodel for evaluating the general toxic effect of biologically active agents and pharmaceuticals
A fragment of cmvEGFP-bGHpolyA with EagI sticky ends containing a green fluorescent protein gene under a cytomegalovirus promoter and terminated by a polyA-sequence of the bull growth hormone gene, cut and purified from a previously created by us plasmid [24], was cloned into a prepared pTZHArbWAР
Summary
The main goals of obtaining genetically modified animals are to change the characteristics of animals, to acquire new opportunities for them, to eliminate some negative qualities in the desired direction for humans. It’s possible to obtain animals with new qualities like resistance to various diseases, increased productivity, changed characteristics of milk, meat, blood, etc. In contrast to the mouse model, the rabbit has a number of advantages. It has been used for studying human diseases for a long time, and it is used as a biomodel for evaluating the general toxic effect of biologically active agents and pharmaceuticals. Rabbits quickly reach sexual maturity, have a short gestation period, and produce large offspring with relatively simple maintenance
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