Erythrocytosis is a rare, acquired or natural disease characterized by an increased number of red blood cells. Congenital erythrocytosis (ECYT1-8) is a group of diseases with a genetic change in one of the genes involved in the process of regulating erythropoiesis. So far, nine genes have been directly linked to congenital erythrocytosis, namely EPOR, VHL, EGLN1, EPO, EPAS1, HBB, HBA1, HBA2 and BPGM. The JAK2 gene in SH2B3 is involved in the development of acquired erythrocytosis, but their role in the development of congenital erythrocytosis remains undefined. In addition, sequential analysis of the coding regions of the genes involved in erythrocytosis leaves 70% of patients with suspected natural erythrocytosis undecided. The purpose of the master's thesis was to determine the genetic differences in the non-coding regions of 11 genes that influence the development of erythrocytosis and to confirm their presence in the genomes of patients. In the first part of the task, we analyzed the non-coding regions of 11 genes with bioinformatics tools. In reviewing the literature, we determined the genomic locations of noncoding regions, histone locations, methylation sites, and genomic locations of CpG islands. The obtained data were compared with the results of NGS analyzes from previous studies on patients and genetic variants that could influence the development of the nature of erythrocytosis were determined. In addition, we analyzed genetic variants determined by tag (ID SNP), association with erythrocytosis, or increased hematocrit, matching with the cryptic exon region and CADD pathogenicity prediction. Of the selected genetic variants, only seven genetic variants could be assigned to the SNP ID. Two of these were associated with elevated hematocrit and elevated EPO expression. Because CADD is lower than 20 in all genetic variants, which is the limit for pathogenicity, most genetic variants were not included in further experimental analysis. In the second part of the task, genetic variants of rs551238 and rs1617640 of the EPO gene, which were previously associated with elevated hematocrit, were included in the experimental analysis. Genotyping was performed on 26 sample DNA samples. The results indicated a possible association between the analyzed SNPs, as the genotypes matched in each sample. Based on the determination of the genotype of one SNP, we were also able to predict the genotype of the other. Analysis of genotypes of family members indicated autosomal dominant inheritance.
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