Since 1970’s gene therapy remains very actual, but still unachievable problem. Discover of genome editing technologies using specific nucleases gives us new possibilities in the treatment of various diseases, including monogenic disorders. This review summarizes brief characteristics of CRISPR/Cas9 method, basic principles, advantages and disadvantages of the method and provides examples of successful application of CRISPR/Cas9 to correct mutations of monogenic hereditary diseases in mouse models and human embryos.

Autism spectrum disorder (ASD) is a developmental disorder of the nervous system characterized by impaired social interaction, communication and stereotypical and repetitive behavior. The aim of this review is to summarize the main results of research conducted in search for genetic causes as well as the disclosure of the major pathogenetic mechanisms of autism.

Methylmalonic aciduria (ММА) - genetically heterogeneous group of hereditary diseases, which characterized by increase of methylmalonic acid (MA) in biological fluids. Biochemically ММА is divided into two groups: isolated and combined. The first one includes diseases associated with mutations in the genes of methylmalonyl-CoA mutase, methylmalonyl-CoA epimerase and diseases caused by disturbance of vitamin B12 metabolism. The second one includes combined MMA with homocysteinemia/homocystinuria caused by mutations in several genes involved in mitochondrial biogenesis. Early diagnosis of the disease is extremely important, since developed approaches to diet therapy of MMA, and the using of hydroxocobalamin for B12-responsive forms allows virtually neutralize all the clinical manifestations of the disease. One of the difficulties of DNA diagnostics is verification of variants in the genes, which pathogenicity is not clear enough. In this research, we studied a biochemical phenotype analysis of 31 patients with isolated MMA. DNA diagnostics was carried out in 18 patients. Mutations in the MUT gene were found in 13 patients. Mutations in the MMAA gene were found in 3 patients. In 2 patients mutations were not found in the MMAA gene. It is planned to study MMAB gene. At first time in our research were found mutations in the MUT gene which were analyzed with programs for the prediction of pathogenicity.

Genetic anomalies: the influence on efficiency of therapy, survival of patients with multiple myeloma, a role in assessment of minimal residual disease. In our research the frequency of occurrence of genetic anomalies and their predictive potential at multiple myeloma is determined. Detection of anomalies was carried out by standard cytogenetic method and fluorescent in situ hybridization (FISH). IgH reorganization (t(11;14), t(4;14), etc.), del(13)(q14), del(17)(p13.1), anomalies of 1 chromosome, hypodiploidy, hyperdiploidy, combined and complex karyotypes decided at patients with multiple myeloma. Genetic anomalies were detected in 57.1% of patients, the frequency of occurrence of anomalies of 1 chromosome, t(11;14), del(13)(q14), t(4;14), del(17)(p13.1), hyperdiploidy, hypodiploidy - in 28.6%, 20.3%, 18.1%, 6.8%, 5.6%, 3.6 and 2.9%, respectively. Duration of observation has averaged 5 years and has shown that del17p13.1 reduced overall survival, t(11;14) didn’t influence the forecast. The complex karyotype has shown negative impact on survival indicators, generally due to existence predictively of adverse anomalies (del(17)(p13.1), del(13)(q14), t(4;14), (+1q)). The Minimal Residual Disease can be revealed by means of flow cytometry and also genetic methods of researches directed to identification of earlier defined anomalies.

Genomic dosages of active ribosomal genes (AcRG) were determined in genomes of 44 apparently healthy subjects (13 men and 31 women aged 17 to 65 years). Previously, polymorphisms of the genes for phase II xenobiotic detoxification enzymes (GXDE) GSTM1 and GSTТ1 were identified in the same subjects. After that, an analysis of combinations of GXDE variants and AcRG copy numbers was performed for each individual genome. The analysis has shown that genomes carrying non-protective (functionally inactive) alleles of GSTM1 and GSTТ1 contain significantly higher genomic dosages of AcRG than genomes carrying functionally active (‘favourable’) alleles of these genes. The findings suggest that the genomic dosage of AcRG exerts certain influence on survival probability (viability) of carriers of the GXDE variants we studied.

Thyroid cancer is the most common endocrine malignancy. The key approach for thyroid cancer diagnosis is cytology of fine needle aspiration biopsy (FNA) samples. FNA specimens have indeterminate cytology in 20-30%. This results in wrong clinical diagnosis and impropriate treatment strategy. Currently known mutations describe vast majority of thyroid cancer cases. Detection of the driver mutations is supposed to improve diagnostic accuracy. The aim of the work is to develop next-generation sequencing based diagnostic panel for thyroid cancer. The analysis of the English-language literature, COSMIC database (DB), as well as results of the research project the Cancer Genome Atlas was performed. In total, 456 point somatic mutations in 25 genes, 23 genetic translocations, and 3 copy number variations (CNV) mutations were identified. Using AmpliSeq Designer, 2 Custom Panels were created - for the detection of point mutations, small indels and CNV (1) and for the detection of translocations (2). The custom Panel for the detection of point mutations, small indels and CNV contains 221 primer pairs in 2 pools, covering 99.59% selected targeted regions. The design also incorporated the regions of the RET gene for detection of germline mutations associated with hereditary medullary thyroid cancer. RNA Gene Fusion designs tool in AmpliSeq Designer was used to design the Panel for the detection of 23 translocations.