Novel strategy for AAV vector design in gene therapy and its biosafety assessment.

To develop a novel AAV vector design for gene therapy by synthesizing the AAVITR microcarrier sequence, constructing and identifying the amplification plasmid, and evaluating the properties of the microcarrier prepared by heat denaturation. We also assess the biosafety of this new vector by examining its toxicity, immunogenicity, and potential for immune cell infiltration. The AAVITR micro carrier sequence was designed and synthesized, followed by the construction of an amplification plasmid to express the vector sequence and the identification of pUC57-minivector. The in vitro expression of the AAVITR micro carrier was analyzed and its safety was assessed through toxicity, immunogenicity, and immune cell infiltration assays. In safety assessments, the new vector demonstrated superior survival and apoptosis rates compared to conventional vectors. Additionally, the AAV-ITR micro carrier showed significant advantages in immune cell infiltration, cytokine levels, and antibody production, indicating a potent immune response. The novel AAVITR micro vector developed in this study offers improved safety and efficacy in gene therapy, with notable advantages in immune response and biosafety, providing a promising alternative to traditional AAV vectors.