Since the very first publications on preimplantation genetic testing, researchers have faced a serious problem — a high mosaicism level in the preimplantation human embryos obtained by means of in vitro fertilization cycles. The nature of this mosaicism and its high impact on embryo development draws attention to this issue. In this research we studied the cells from different parts of preimplantation human embryos with mosaicism in the trophectoderm cells detected using Next-generation Sequencing (NGS). Six human blastocysts with mosaicism in their trophectoderm cells were each sectioned in three parts: two containing only trophectoderm cells and one predominantly inner cell mass. These parts were then analyzed individually. Our data indicate that the proportion of aneuploid cells in bioptate taken for preimplantation genetic testing does not necessarily reflect the true chromosomal status of the whole embryo and cannot be extrapolated to that in the embryoblast cells. The results of our study strongly suggest that mosaicism revealed in blastocyst reduces the likelihood of finding the euploid chromosome set in the other parts of the embryo. Karyotypes of cells from different parts of mosaic embryos show low concordance. Chromosomal abnormalities in mosaic embryos are unpredictably diverse, which may lead not only to loss of conception, but also to the development of genetic disease in the offspring. According to our data, the mosaic rate tends to increase in the samples containing trophectoderm adjacent to the embryoblast, which may have physiological significance for the implantation. Comparative studies focused on the concordance of mosaicism level of and the type of chromosomal abnormalities detected in different parts of preimplantation human embryos will improve clinical recommendations regarding the transfer of mosaic embryos. Since the very first publications on preimplantation genetic testing, researchers have faced a serious problem — a high mosaicism level in the preimplantation human embryos obtained by means of in vitro fertilization cycles. The nature of this mosaicism and its high impact on embryo development draws attention to this issue. In this research we studied the cells from different parts of preimplantation human embryos with mosaicism in the trophectoderm cells detected using Next-generation Sequencing (NGS). Six human blastocysts with mosaicism in their trophectoderm cells were each sectioned in three parts: two containing only trophectoderm cells and one predominantly inner cell mass. These parts were then analyzed individually. Our data indicate that the proportion of aneuploid cells in bioptate taken for preimplantation genetic testing does not necessarily reflect the true chromosomal status of the whole embryo and cannot be extrapolated to that in the embryoblast cells. The results of our study strongly suggest that mosaicism revealed in blastocyst reduces the likelihood of finding the euploid chromosome set in the other parts of the embryo. Karyotypes of cells from different parts of mosaic embryos show low concordance. Chromosomal abnormalities in mosaic embryos are unpredictably diverse, which may lead not only to loss of conception, but also to the development of genetic disease in the offspring. According to our data, the mosaic rate tends to increase in the samples containing trophectoderm adjacent to the embryoblast, which may have physiological significance for the implantation. Comparative studies focused on the concordance of mosaicism level of and the type of chromosomal abnormalities detected in different parts of preimplantation human embryos will improve clinical recommendations regarding the transfer of mosaic embryos.

The phenomenon of evolutionary fixation of agrobacterial sequences (cT-DNA or cellular transferred DNA) in plant genomes is well known in nature. It was previously considered, that all of cT-DNA-containing species, except Linaria vulgaris , have multiple inverted cT-DNA repeats. Deep studying of general features of cT-DNA brings us closer to understanding the causes and mechanisms of its fixation in plants genomes. We combined multiple long-range PCR with genome walking for studying extended structure of cT-DNA. Using digital PCR method, we estimated copy number of cT-DNA elements. NGS with low covering allows us to develop a set of microsatellite markers, also used for copy number estimation. According to new data, cT-DNA elements in L. vulgaris form an inverted complex repeat of two simple direct repeats. After cT-DNA integration, cT-DNA sequence duplication events took place at least two times. The phenomenon of concerted evolution of cT-DNA sequences as well as some details of this process have been shown for the first time. We have shown, that L. vulgaris, as well as other cT-DNA containing species, has inverted structure of repeats. This fact indicates possible existence of some general causes and mechanisms of cT-DNA fixation in plant genomes during evolution.

Ribosomal RNA (18S, 5.8S, 28S) gene clusters in genomes form regions that consist of multiple tandem repeats. They are located on a single or several pairs of chromosomes and play an important role in the formation of the nucleolus responsible for the assembly of ribosome subunits. The rRNA gene cluster sequences are widely used for taxonomic studies, however at present, complete information on the avian rDNA repeat unit structure including intergenic spacer sequence is available only for the chicken (Gallus gallus domesticus Linnaeus, 1758). The GC enrichment and high-order repeats peculiarities within the intergenic spacer described for the chicken rDNA cluster may be responsible for these failures. The karyotype of the Japanese quail (Coturnix japonica Temminck et Schlegel, 1849) deserves close attention because, unlike most birds, it has three pairs of nucleolar organizer bearing chromosomes, two of which are microchromosomes enriched in repeating elements and heterochromatin that carry translocated terminal nucleolar organizers. Here we assembled and annotated the complete Japanese quail ribosomal gene cluster sequence of 21166 base pairs (GenBank under the registration tag BankIt2509210 CoturnixOK523374). This is the second deciphered avian rDNA cluster after the chicken. Despite the revealed high similarity with the chicken corresponding sequence, it has a number of specific features, which include a slightly lower degree of GC content and the presence of bendable elements in the content of both the transcribed spacer I and the non-transcribed intergenic spacer.