Pv11 cell culture
One of our research objectives is Pv11 cell culture, derived from chironomid Polipedilum vanderplanki. Similarly to the source organism, these cells able to revive after complete desiccation.
Current projects within this research area:
- identification of chromosome rearragements and "fragile" genome sites accumulating mutations in Pv11 cells under desiccation conditions;
- genome editing of Pv11, including a use of CRISPR/Cas9 technology;
- expression of some Polypedilum vanderplanki genes in "ordinary" insect cell culture SF9; we study properties of corresponding proteins and try to improve stress tolerance of Sf9 culture;
- identification of ultrastructure changes in Pv11 cells during desiccation and rehydration.
Methyltransferases preventing the aging of proteins
The genome of African chironomid P.vanderplanki contains a specific gene clusters, which are formed by numerous paralogues. These genes code important proteins for successful transition to the anhydrobiotic stage. One of the such clusters consist of 14 L-isoaspartyl methyltransferases (PIMT) – the object of our current research. At the present stage we have expression data for these genes from the experiments of desiccation-rehydration of the larvae. There is a strong response in case of almost all PIMT genes, except classic one, which is not included into the cluster. We purified recombinant proteins and estimated the activity level at normal conditions for human PIMT using standard substrate – DSIP peptide with L-isoaspartyl residue in the middle. In these conditions only classic enzyme and one more from the cluster PIMT2 showed presence of the methyltransferase activity. At this time, we analyze CAGE data for Pv11 cell culture and continue experiments with recombinant proteins.
In addition, recently we started to analyze the data from space experiment with Zebrafish.
Etnospecific features of tumor genetic markers within population of Tatarstan Republic
The study of the human transcriptome, patient with lung cancer, namely adenocarcinoma. The main objective of this research is identification of reliable tumor markers associated with the activity of receptors and ligands of PD and PDL1 etnospetsific characteristics of the population of Tatarstan.
Preparation of genomic libraries from clinical blood samples from more than 100 women had breast cancer with of hereditary genetic component and an unclear status of the structure oncogene BRCA1 and BRCA2. For research and clinical purposes design panels, containing BRCA1 and BRCA2 genes, as well as the regulatory regions of the genome associated with these genes and a number of genes functionally associated with the BRCA1 and BRCA2 were made based on the literature data and the results of experiments. The panel will be used in the project initiative the Republic Clinical Oncological Hospital for the early detection of risk groups in the country.
Bacterial metagenom of anhydrobiotoc chironomids Polypedilum vanderplanki
The study of the taxonomic diversity of microorganisms (metagenome) associated with the African chironomid Polypedium vanderplanki, at different stages of individual development. It carried out by decoding the nucleotide sequence of 16S rRNA genes exhibiting a high species specificity. Currently we spend preparation stage of DNA libraries of 16S rRNA genes for subsequent sequencing. The same library is prepared for another model object – sea cancer of Semibalanus genus, specimens of which were taken from different areas of the White Sea. Work is carried out with the genomic DNA extracted from the model objects.
Molecular basis of anhydrobiosis: comparative and evolutionary aspect
The study of molecular-genetic mechanisms anhydrobiosis in the evolutionary series of anhydrobiotic organisms, from prokaryotes to highly organized invertebrates, in particular, for typical anhydrobiont – larvae of chironomid Polypedium vanderplanki (Insecta, Diptera). The purpose is to carry out a comparative analysis of molecular genetic and biochemical and physiological mechanisms of the anhydrobiosis state – anabiotic life in the almost complete absence of water inside the cells and tissues. It is supposed to identify a common set of features and genetic markers necessary for the survival of organisms in the absence of water, i.e. a minimal "survival kit" of anhydrobionts. The work is theoretical, at now for its results a review article is preparing.