The Use of Reproductive Technologies to Produce Transgenic Goats

Recombinant DNA technology has revolutionized the production of therapeutic proteins. Thus, genes of a great number of human proteins have already been identified and cloned. The use of farm animals as bioreactors may be the better choice to produce recombinant therapeutic proteins. For this activity, the term “pharming” was created, referring to the use of genetic engineering to obtain a transgenic or genetically modified animal. Considering the advantages and disadvantages of livestock species, goats appear as a very good model. In addition, the first human commercially approved biological drug (antithrombin (AT)) was produced from the milk of transgenic goats. The aim of this chapter is to present various reproductive technologies used to obtain transgenic goats secreting recombinant proteins in milk. Initially, this chapter presents the methods for embryo production (in vivo and in vitro) to realize the DNA microinjection in pronuclear embryos. Thus, the techniques of superovulation of donors (in vivo embryo production) and ovarian stimulation for oocyte recovery (in vitro embryo production) are described. Also, the methods for DNA microinjection and embryo transfer are detailed in this chapter. Finally, this chapter describes the reproductive procedures used for obtaining transgenic goats by cloning

International Comparisons: Issues of Methodology and Practice

The article discusses the methodology and organization of statistical observation of the level of countries’ economic development. The theoretical basis of international comparisons is singled out and on its basis the comparative evaluation of inconsistency of theoretical positions and the reasons of differences of GDP growth is carried out. Based on the complexity of the formation of homogeneous data sets in order to obtain correct comparison results, a general scheme for the relationship between the theoretical base of international comparisons and PPP constraints is defined. The possibility of obtaining a single measurement of the indicators of national economies based on the existing sampling errors, measurement uncertainties and classification errors is considered. The emphasis is placed on combining the work using the ICP and CPI with the aim of achieving comparability of data in the territorial and temporal cross-section. Using the basic characteristics of sustainable economic growth, long-term prospects for changing the ranking positions of countries with different levels of income are determined. It is shown that the clarity and unambiguity of the theoretical provisions is the defining condition for the further process of data collection and formation of correct analytical conclusions

Conceptual scheme of management of expenditures of an industrial enterprise Концептуальная схема управления затратами промышленного предприятия

The uses analysis of approaches and principles of management of expenditures to identify a general scheme of management, which would allow marking out both the information base of the study and the sequence of interconnected actions on formation of the conceptual scheme of management of expenditures of an enterprise. Account of conditions of uncertainty for a point, interval and area of break-even through the system of balanced indicators gives a possibility to account multidirectionality of possible actions on selection of optimal methods of management of expenditures of an enterprise.В статье на основе анализа подходов и принципов управления затратами определена общая схема управления, которая позволит выделить как информационную базу исследования, так и последовательность взаимосвязанных действий по формированию концептуальной схемы управления затратами предприятия. Учет условий неопределенности для точки, интервала и площади безубыточности через систему сбалансированных показателей дает возможность учесть разновекторность возможных действий по выбору оптимальных способов управления затратами предприятия

Generation of megabase-scale deletions, inversions and duplications involving the Contactin-6 gene in mice by CRISPR/Cas9 technology

Abstract Background Copy Number Variation (CNV) of the human CNTN6 gene (encoding the contactin-6 protein), caused by deletions or duplications, is responsible for severe neurodevelopmental impairments, often in combination with facial dysmorphias. Conversely, deleterious point mutations of this gene do not show any clinical phenotypes. The aim of this study is to generate mice carrying large deletions, duplications and inversions involving the Cntn6 gene as a new experimental model to study CNV of the human CNTN6 locus. Results To generate large chromosomal rearrangements on mouse chromosome 6, we applied CRISPR/Cas9 technology in zygotes. Two guide RNAs (gRNAs) (flanking a DNA fragment of 1137 Mb) together with Cas9 mRNA and single-stranded DNA oligonucleotides (ssODN) were microinjected into the cytoplasm of 599 zygotes of F1 (C57BL x CBA) mice, and 256 of them were transplanted into oviducts of CD-1 females. As a result, we observed the birth of 41 viable F0 offspring. Genotyping of these mice was performed by PCR analysis and sequencing of PCR products. Among the 41 F0 offspring, we identified seven mice with deletions, two animals carrying duplications of the gene and four carrying inversions. Interestingly, two F0 offspring had both deletions and duplications. It is important to note that while three of seven deletion carriers showed expected sequences at the new joint sites, in another three, we identified an absence of 1–10 nucleotides at the CRISPR/Cas9 cut sites, and in one animal, 103 bp were missing, presumably due to error-prone non-homologous end joining. In addition, we detected the absence of 5 and 13 nucleotides at these sites in two F0 duplication carriers. Similar sequence changes at CRISPR/Cas9 cut sites were observed at the right and left boundaries of inversions. Thus, megabase-scale deletions, duplications and inversions were identified in 11 F0 offspring among 41 analyzed, i.e., approximately 25% efficiency. All genetically modified F0 offspring were viable and able to transmit these large chromosomal rearrangements to the next generation. Conclusions Using CRISPR/Cas9 technology, we created mice carrying megabase-scale deletions, duplications, and inversions involving the full-sized Cntn6 gene. These mice became founders of new mouse lines, which may be more appropriate experimental models of CNV in the human 3p26.3 region than Сntn6 knockout mice

Genetic modification of mammalian genome at chromosome level

The review is concerned with a progress in genetic modification of a mammalian genome in vitro and in vivo at chromosomal level. Recently three new approaches for the chromosome biotechnology have been developed: Using Cre/loxP-system a researcher is able to produce targeted rearrangements of whole chromosomes or their segments or particular genes within the genome, and therefore to modify the set, position and copy number of the endogenous elements of the genome. Mammalian artificial chromosomes (MACs) provide a possibility to introduce into genome relatively large segments of alien chromosome material, either artificially constructed or derived from the genome of different species. Using ES-somatic cell hybrids allows to transfer whole chromosomes or their fragments between different genomes within and between species. Advantages and limitations of these approaches are discussed

DEVELOPMENT OF THE GRADUATES’ EMPLOYMENT MONITORING SYSTEM

The article suggests a complex approach to the monitoring of graduates’ employment. Analytical materials about employment of graduates of higher education institutions were formed in accordance with the findings of monitoring