Exogenous Expression of Receptors for AAV9 to Increase the Cell Line Sensitivity to Viral Particle Transduction

The adeno-associated virus serotype 9 (AAV9) vector is currently the leading platform for delivering genes for treating various human diseases. The structural characteristics and properties of AAV9 make it a promising candidate for gene therapy due to its unique qualities. AAV9 has a high affinity for muscle tissues, including the heart muscle, and can effectively cross the blood-brain barrier. This makes it a valuable tool for treating diseases associated with the central nervous system and muscular and cardiac conditions. Despite the results demonstrated by this vector in in vivo experiments, where it exhibits high efficiency in cell transduction, AAV9 poorly transduces cell lines in vitro. This has made it difficult to further develop this promising vector. To solve the presented problem within the framework of this thesis, one approach to solving the problem was used. The aim of the study is to enhance the sensitivity of the HEK293T cell line to AAV9 transduction through the exogenous expression of viral particle receptors in cells. As part of the research, the efficiency of AAV9 transduction was assessed in comparison with other serotypes of AAV on the HEK293T cell line, based on a literature analysis, potential receptors that could increase AAV9 transduction were identified. Their genes were cloned into vectors with a reporter system. After overexpression in cells, the receptors that increased AAV9 transduction in the HEK293T line were selected under conditions of transient expression. The study identified KIAA0319L as a receptor that significantly increased AAV9 transduction levels in HEK293T cells when expressed exogenously. The data obtained can serve as a basis for further research into the effect of KIAA0319L overexpression on AAV9 transduction in other cell lines. For instance, in the mouse myoblast line C2C12, which is widely used to test genetic treatments for muscle diseases, and further create stable cell lines highly expressing this receptor for effective AAV9 transduction.