Molecular biology and genetics
This course deals with the fundamental principles governing the flow of genetic information, from DNA replication to regulation of gene expression. It explores the intricacies of DNA biosynthesis and repair, as well as the processes of DNA transcription, and translation of messenger RNA. The course covers the regulation of gene expression in both prokaryotes and eukaryotes, including the regulation of eukaryotic cell division and differentiation. Practical methodologies such as polymerase chain reaction, molecular cloning, and next-generation sequencing will also be examined. Additionally, the course addresses the genetic basis of complex traits, inborn errors of metabolism, and mechanisms of sex determination, while providing the theoretical foundation for detecting hereditary pathologies and infectious diseases, genome editing, and generating genetically modified organisms. Furthermore, it covers the molecular mechanisms involved in the adaptation of living organisms to adverse environments, particularly oxidative stress. This comprehensive approach equips students with a robust understanding of molecular biology and genetics, preparing them for advanced research or clinical applications.
The main dogma of molecular biology. The flow of genetic information in pro- and eukaryotes and the basic principles of its regulation.
DNA replication — initiation, elongation, termination. Formation of phosphodiester bonds. DNA replication in E. coli. DNA replication in eukaryotes. Replication at the ends of linear chromosomes.
Terminology. Repair in E. coli by excision. Repair during replication. Reversal of damages. SOS repair.
Initiation and elongation. Termination. RNA polymerases of pro-, eukaryotes and archaebacteria. RNA processing. Ribozymes.
Activation of amino acids. Initiation, elongation, and termination of protein biosynthesis. Regulation of translation.
Genetic code. Genotype and phenotype. Biochemical bases of heredity. Model organisms in genetics. Recombination and chromosomal crossing over. Cytoplasmic inheritance. Epigenetic factors.
General principles. Repression and induction. Operons and regulons. Peculiarities of gene expression in pro- and eukaryotes. Positive and negative control. Lactose and tryptophan operons. Catabolite repression. DNA protection.
Regulation of quorum sensing in bacteria. Regulation of chemotaxis in bacteria. Factors that regulate the transition from anaerobic to aerobic metabolism in Escherichia coli. Regulation of nutrient assimilation in bacteria. Bacterial chaperones.
Response to nutrient availability in baker’s yeast. Conceptions regarding proteasome and ubiquitination. Regulation of iron metabolism. Iron regulatory proteins. Regulation of hypoxia response in eukaryotes. Insulin signaling pathway and regulation of cholesterol biosynthesis. Regulation of circadian rhythms.
Active forms of oxygen in biological systems. Oxidative/reductive stress. Bacterial regulons OxyR and SoxRS. Yap1 stimulon in yeasts. The Keap1/Nrf2 system in animals.
Signaling pathway, which is regulated by the Notch receptor. The role of the Wnt pathway in morphogenesis. AP-1 factor; Jun and Fos proteins. Morphogens Dpp, BMP, and Hedgehog. Cell reprogramming; Yamanaka factors.
Classification of gene mutations. Methods of their research. Mechanisms and causes of gene mutations. Typical pedigree profiles for different types of inheritance. Reasons for deviations from typical profiles. Penetration. Expressiveness. Interaction of genes. Expansion diseases. Medscape, Cochrane Library, OMIM, Orphanet.
Genomic mutations. Mixoploids: mosaicism and chimerism, mechanisms of their appearance. Mechanisms of aneuploidy. Chromosomal syndromes caused by aneuploidy: Turner, Klinefelter, triple X, Down, Edwards, Patau. Risk factors for chromosomal mutations.
Deletions, duplications, inversions, translocations, ring chromosomes, isochromosomes. Chromosomal syndromes caused by chromosomal rearrangements: "Cri du chat", Williams, DiGeorge, Prader-Willi, and Angelman syndromes; the phenomenon of genomic imprinting.
The main sources of clinical data: Medscape, Cochrane Library, OMIM, Orphanet. Mechanisms of inheritance: autosomal recessive, autosomal dominant, X linked dominant and recessive. X-linked diseases in women; irregular lyonization of the X chromosome and its consequences.
Twin studies, genealogical, cytogenetic, biochemical, and molecular methods.
Polymerase chain reaction (PCR) principle. The main types of PCR. Identification of heritable pathologies and infectious disease by PCR method. The basics of primer design. Sanger sequencing.
“Anatomy” of a cloning vector. The use of restriction enzymes. Transformation. Ligation. Expression systems. Tags for recombinant proteins. Contemporary concepts regarding gene editing. Concepts regarding gene therapy.
Metabolism, genes, and enzymes. Classification of inborn errors of metabolism. Clinical signs. Principles of diagnosis. The method of tandem mass spectrometry (TMS). Phenylketonuria (PKU), description and frequency of newborns with the disease. Gene and product characteristics. Mechanism of development of symptoms. Method of treatment. The importance of early diagnosis. Neonatal screening.
Chromosomal mechanism of sex determination. X and Y human chromosomes. SRY locus. Molecular and hormonal mechanism of sex determination. Gonadal dysgenesis. Androgen insensitivity syndrome. Swyer syndrome. Congenital adrenal hyperplasia. Disorders of sex development (DSDs). Intersex people.
NGS workflow. Bioinformatics analysis of NGS data. Commonly used tools for NGS data analysis. Modern nomenclature of NGS results. Criteria for NGS interpretation, prediction algorithms, and necessarily databases. Clinical application of NGS.
GWAS analysis. Genetics and environmental factors measurement. Genome-Wide Association Study (GWAS) - method description. GWAS catalog. Interpretation of genomic study; definition of absolute risk, relative risk, odds ratio. Examples of GWAS success (diabetes, hypertension, schizophrenia, cancer). Genome study value in clinical practice.
