Dear Visitor, we can set up here your advertisements upon your wish. Please get in touch with us.
  Login:    Sitemap:    Search:

grant:KP-06-H63/1/13.12.2022, Comparative analysis of the effectiveness of new antibacterial agents based on different types of antisense oligonucleotides using different molecular mechanisms of RNA inhibition awarded to Prof. Robert Penchovsky by the Bulgarian National Science Fund (BNSF)

The project "Comparative analysis of the effectiveness of new antibacterial agents based on different types of antisense oligonucleotides using different molecular mechanisms of RNA inhibition" is a continuation of long-term research in the field of synthetic biology and the development of new antibacterial agents to be tested and optimized, through appropriate modifications or different molecular mechanisms. The planned fundamental experiments include applying the theoretically possible methods for optimizing the effectiveness of antibacterial agents through modifications in their structure and the application of various molecular mechanisms of RNA inhibition. The main focus of our project is the well-known antisense oligonucleotides to our team, with which we have been actively working for the last decade in the laboratory of Prof. Penchovski at the Faculty of Biology of Sofia University "St. Kliment Ohridski” as a continuation of his experience from his work with bacterial riboswitches at Yale University, USA, together with Ronald Breaker (discoverer of riboswitches). Antisense oligonucleotides are agents that bind specifically to the molecule they target. There are currently three generations of antisense oligonucleotides, each with its characteristics, advantages, and limitations. For our project, we will design different antisense oligonucleotides that will be representatives of different generations, as well as those that will be chimeric. We will apply them in bacterial cells to perform genetic control through their specific binding to bacterial riboswitches. Bacterial riboswitches are structural RNA domains, most commonly located in the 5'-untranslated region of messenger RNAs that specifically recognize and bind metabolites. To date, no evidence of riboswitches has been found in the human genome, making bacterial riboswitches potentially suitable targets for developing antibacterial agents. The structure of most riboswitches is known, and in international bioinformatics databases: GenBank, Rfam, including in the RSwitch database created by us the sequences of individual species and their representatives are also available. As a result, specific binding of antisense oligonucleotides and riboswitches results in conformational changes that enable messenger RNAs to regulate their expression without needing regulatory proteins. This leads to the impossibility of vital metabolites for the bacteria to be synthesized by the cells and/or transported inside it from the external environment, leading to a bacteriostatic or bactericidal effect.

The comparison of individual variants of antisense oligonucleotides will allow us to analyze them and, as a result, answer several open questions for the scientific community regarding the design of antisense oligonucleotides - their basic nucleotide skeleton, the generation to which they belong, and the possible modifications of the structure them As a separate task within the project, we aim to expand our knowledge of the types of cell-penetrating peptides and their behavior when loaded with antisense oligonucleotides by conducting experiments with selected antisense oligonucleotides and different cell-penetrating peptides. As a result of the experiments, we will observe the influence on the effectiveness of individual complexes of cell-penetrating peptides and antisense oligonucleotides. We will compare the data obtained and give a conclusion about the effectiveness and applicability of each of them. This newly obtained information will be helpful in the further studies of colleagues around the world. Through these data, we will propose an efficiency ranking and reduce inefficient attempts to import an antisense oligonucleotide into a bacterium, the time to create new therapeutic agents, and the cost of experiments and personnel. After completing our highly successful project "Design and experimental validation of chimeric antisense oligonucleotides as antibacterial agents", the results of which were published in 20 scientific publications with a total impact factor of 84 points (Q1-4 = 274 and 86 citations) and we created 16 software products available on the official website of Prof. Robert Penchovsky, we are confident that with the creation of new antisense oligonucleotides linked to different cel-penetrating peptides, based on different molecular mechanisms of RNA inhibition, to compare their effectiveness, we will obtain fundamental data through which we will give new directions and draw new horizons, accelerating the process of creating antibacterial agents against serious, life-threatening pathogenic bacteria. The research of the project funded with 102 300 EUR.