Moscow State University, Faculty of Biology, Department of Genetics
4
Scopus Publications
Scopus Publications
Multi-omics study of molecular and genetic bases of orthostatic hypotension Elena Zelenova, Veronika Daniel, Maria Bruttan, Lilya Artemieva, Ekaterina Spektor, Aleksandra Mamchur, Daria Kashtanova, Mikhail Ivanov, Gerel Abushinova, Lorena Matkava, Antonina Rumyantseva, Aleksey Ivashechkin, Pavel Grebnev, Liliya Golubnikova, Sergey Mitrofanov, Anna Akopyan, Olga Beloshevskaya, Tatiana Saliyeva, Irina Tarasova, Irina Strazhesko, Vladimir Yudin, Valentin Makarov, Anton Keskinov, Olga Tkacheva, Sergey Yudin, Veronika Skvortsova Clinical Epigenetics, 2025 Orthostatic hypotension is a sharp decrease in blood pressure when an individual transitions from a supine to an upright position. OH affects at least 30% of older adults. It is attributed to the dysfunction of the autonomic innervation and decreased vascular bed capacity. Genomic (n = 2526), methylomic (n = 910), and transcriptomic (n = 391) data from centenarians aged 90 years and older were used to examine molecular and genetic factors for OH. No statistically significant genetic predictors of OH were identified. However, the study revealed numerous epigenetic markers of OH indicative of general aging, such as DNA hypomethylation. The predictive DNA methylation-based model for orthostatic hypotension demonstrated an average accuracy of 79%. The transcriptome analyses highlighted associations between OH and inflammation pathways, as well as other age-related biological processes. Integrated omics and clinical data have identified six key mechanisms associated with orthostatic hypotension: metabolic dysregulation, impaired muscle tone, altered cell proliferation, inflammation, humoral regulation, and neural regulation.
Visualizing Nucleic Acid Loci with the CRISPR-Cas System: Current Approaches Gerel A. Abushinova, Victoria V. Zherdeva, Ekaterina M. Vassina, Liliya G. Maloshenok Journal of Biomedical Photonics and Engineering, 2024 The visualization of genomic loci in living cells is crucial for detecting mutations and observing the spatial proximity of DNA regions in the 3D nuclear environment. One of the primary applications of the clustered regularly interspaced short palindromic repeats–CRISPR-associated protein 9 (CRISPR/Cas9) system is the non-invasive, real-time labeling of DNA loci in living cells, enabled by its unique characteristics. Visualization of genomic loci has been made possible by the use of an endonuclease-inactive form of the Cas9 protein (dCas9) and sgRNA in combination with fluorescent molecules. However, using CRISPR/Cas9 for targeting DNA regions has certain limitations, the most significant being suboptimal signal-to-noise ratio, the need for multiplexed labeling, and the large size of the Cas fluorescent reporter sytem, which impacts the complex’s functionality and complicates its delivery. Current variations of the method using CRISPR/dCas9 overcome these limitations in different ways. This review examines the evolution of genome locus visualization methods based on CRISPR/Cas9 from the initial use of the system to the present.
Tet-Regulated Expression and Optical Clearing for In Vivo Visualization of Genetically Encoded Chimeric dCas9/Fluorescent Protein Probes Liliya Maloshenok, Gerel Abushinova, Natalia Kazachkina, Alexei Bogdanov, Victoria Zherdeva Materials, 2023 The catalytically inactive mutant of Cas9 (dCas9) endonuclease has multiple biomedical applications, with the most useful being the activation/repression of transcription. dCas9 family members are also emerging as potential experimental tools for gene mapping at the level of individual live cells and intact tissue. We performed initial testing on a set of tools for Cas9-mediated visualization of nuclear compartments. We investigated doxycycline (Dox)-inducible (Tet-On) intracellular distribution of constructs encoding dCas9 orthologs from St. thermophilus (St) and N. meningitides (Nm) fused with EGFP and mCherry fluorescent proteins (FP) in human A549 cells. We also studied time-dependent expression of these chimeric fluorescent constructs (dCas9-FP) after Tet-On induction in live cells and compared it with the time course of dCas9-FP expression in experimental dCas9-FP-expressing tumor xenografts using a combination of fluorescence imaging and in vivo contrast-assisted magnetic resonance imaging for assessing the extent of tumor perfusion. In vivo Dox-induction of mCherry-chimera expression occurred in tumor xenografts as early as 24 h post-induction and was visualized by using optical clearing (OC) of the skin. OC via topical application of gadobutrol enabled high-contrast imaging of FP expression in tumor xenografts due to a 1.1–1.2-fold increase in FI in both the red and green channels.
Visualizing the Nucleome Using the CRISPR–Cas9 System: From in vitro to in vivo Liliya G. Maloshenok, Gerel A. Abushinova, Alexandra Yu. Ryazanova, Sergey A. Bruskin, Victoria V. Zherdeva Biochemistry Moscow, 2023 One of the latest methods in modern molecular biology is labeling genomic loci in living cells using fluorescently labeled Cas protein. The NIH Foundation has made the mapping of the 4D nucleome (the three-dimensional nucleome on a timescale) a priority in the studies aimed to improve our understanding of chromatin organization. Fluorescent methods based on CRISPR-Cas are a significant step forward in visualization of genomic loci in living cells. This approach can be used for studying epigenetics, cell cycle, cellular response to external stimuli, rearrangements during malignant cell transformation, such as chromosomal translocations or damage, as well as for genome editing. In this review, we focused on the application of CRISPR-Cas fluorescence technologies as components of multimodal imaging methods for in vivo mapping of chromosomal loci, in particular, attribution of fluorescence signal to morphological and anatomical structures in a living organism. The review discusses the approaches to the highly sensitive, high-precision labeling of CRISPR-Cas components, delivery of genetically engineered constructs into cells and tissues, and promising methods for molecular imaging.
