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Transgenesis through the spermatogonia stem cells in the animals and prospects of its application in Bilotserkiva NAU
Genetic modification and transgenesis are based on the technology of obtaining recombinant DNA based on. Transgenesis is an artificial transfer of a gene (or DNA fragment) from one organism to another. Genetic modification can occurs when the DNA sequence in an organism is modified without the transfer of foreign genes.
Living organisms with artificially modified genome are called genetically modified (GM) or transgenic. Transgenic animals are created for the following main purposes:
- Basic biological research (rodents);
- Modeling of specific human disease for biomedical applications (rodents and large animals);
- In vivo production of therapeutic proteins;
- Enhancement of useful traits in agriculturally important animals.
The first transgenic animal (mouse) was obtained in 1981 as a result of the collaboration of several scientific laboratories in the United States.
Today there are several widely used methods of introducing of foreign DNA into the animal's genome, for the production of transgenic animals:
- microinjection method;
- the use of modified embryonic stem cells followed by the transfer of transformed into early embryo;
- use of spermatogonal stem cells and sperm as a vector for DNA transfer.
Microinjection of recombinant DNA into fertilized oocytes remains the most popular method of introduction of foreign genes into the genome of animals. This method requires highly qualified and experienced specialists, as well as expensive equipment and reagents. Another method to create a transgenic mice used today is transgenesis through modification of embryonic stem cells followed by introduction of modified cells into early donor embryo.
We emphasize that the methods described above have been well developed and are widely used today for the production of transgenic mice. Therefore, the market for transgenic mice is now sufficiently saturated. However, these methods are way less effective for rats and other species. The number of transgenic mice models is calculated by tens of thousands, while the number of available transgenic rats does not exceed 500 models. Rat, as a model, surpasses the mouse as it is closer to the human biology. The access to new and desired models of rats will give scientists in many research areas the opportunity to choose experimental animals that are closer to man.
According to “Marked and Marked” report “The rat model market is estimated at USD 358.5 Million in 2014 and is expected to reach USD 602.3 Million by 2020, growing at a CAGR (Combined Average Growth Rate) of 9.4% from 2015 to 2020”.
Advancements in gene editing technologies such as CRISPR/Cas 9, TALEN, Zinc Finger Nucleases make generation of rats with some types of genetic modifications technically possible and economically reasonable.
Worldwide, the spermatogonia mediated transgenic technology is being developed at several academic institutions in the United States, Canada and Japan. Currently, commercial companies cannot use this technology because of its novelty and the absence of trained specialists in this field. Some steps utilized in this technology require specific knowledge, skills and experience. The proposed laboratory of genetic zoinengineering will be a biotechnological unit of the Bila Tserkva National Agrarian University for the production of genetically modified animals. It is assumed that the laboratory will be involved in:
1. Implementation of a reliable technological process for the production of transgenic model animals;
2. Proposition for the creation of genetically engineered animal models for use in scientific research by scientists of BNAU;
3. Having successful examples of the creation and use of models animals, the laboratory will offer services for the generation of transgenic model animals at the national and international levels;
4. Extension of the technology onto other species of animals, where generation of genetic modifications by traditional methods (oocyte or embryo manipulation) have not been developed, is not effective, or has an excessively high cost. Creating the first modification in guinea pig is a priority task;
5. Creation and supply of genetically modified guinea pigs of high commercial value for general use in fundamental study of infectious diseases, allergies, immunity and asthma;
In the future, the laboratory will aim on creation of genetically improved farm animals with increased productivity or resistance to diseases, as well as animals for the biological production of proteins of pharmacological significance.
The laboratory will become the first scientific-experimental unit in Ukraine to offer models of genetically engineered laboratory animals created by alternative and more effective transgenic technology.
Key words: genetic engineering, transgenosis, recombinant DNA, transgenic animals, spermatogonial stem cells, transplantation of spermatogonium.
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