Kitosan hidrojeline gömülü ov-GnRH mRNA aşisinin geliştirilmesi

Physical castration has been practiced in many mammalian species for centuries for various reasons, including preventing uncontrolled reproduction in farm animals, suppressing aggressive behavior, improving meat quality, and preventing uncontrolled reproduction in domesticated, stray, and wild animals, as well as in zoos and wildlife parks. Because physical castration is not a preferred practice, immunizing mammals against Gonadotropin-Releasing Hormone (GnRH) to suppress their reproductive functions (GnRH immunization-immunological castration- immunocastration) has emerged as a biotechnological solution. Protein and DNA vaccines developed for immunocastration are available in the literature. mRNA vaccines, a new technology, are considered a potential candidate to address the technological shortcomings of commercially available protein-based vaccines due to their superior features, including i) faster, higher quality, and easier production in a laboratory environment, ii) production of the encoded antigen in its native form (through in situ posttranslational modifications), and iii) the induction of a long-lasting, healthy immune response to the in situ produced antigen. There is no mention of an mRNA-based immunocastration vaccine, or more specifically, an mRNA-based GnRH vaccine, in the literature. This study aimed to synthesize Ov-GnRH mRNA in vitro using the Ovalbumin-GnRH (Ov-GnRH) gene region, whose immunocastration success has been previously demonstrated in different species. In this context, the DNA region encoding Ov-GnRH was rearranged as a template, and Ov-GnRH mRNAs capable of high-yield expression in the host were synthesized. To eliminate the need for booster doses in traditional immunization practices and to provide long-term and effective immunization with a single injection, Ov-GnRH mRNAs were loaded into a thermosensitive chitosan hydrogel, and characterized in vitro condition. To achieve this goal, a thermosensitive chitosan hydrogel was prepared that, when injected into a living organism, gels at body temperature and provides a sustained/slow release of the loaded Ov-GnRH mRNA. The synthesized biomaterial's mRNA loading capacity, mRNA release kinetics, dissolution profile under physiological conditions, and cytotoxicity were investigated in vitro.