Implementing the human artificial chromosome gene therapy platform remains challenging, but continuous animal model research will advance the platform closer to clinical trials

Abstract

A normal degree of ectopic gene expression, infinite retention in target cells without chromosomal integration, minimal risk of cell or neoplastic transformation, and minimal or no immunogenicity are all critical characteristics for vectors employed in gene therapy. HACs were produced and used as autonomous vectors to compensate for genetic defects in mouse and human cell cultures. Bottom-up human artificial chromosomes (HACs) were studied for functional transgene expression in vitro and in vivo mice models. The primary advantages of synthesized alphoid-HACs over top-down HACs are their defined and documented structure, as well as their relative simplicity of modification in adding numerous Cre-lox-type transgen loading sites. The HAC transfer method's efficacy has greatly increased in recent years. Despite significant progress in developing alphoid-HAC-based gene therapy models, the technology still has a number of drawbacks, including low HAC efficiency, complex repeated HAC alphoid-DNA structure, large DNA fragmentation difficulties outside eukaryotic cells, inefficient transfer of chromosomes to target cells, and variable mitotic stability. The quantity and quality of PSC-derived or reversibly immortalized stem/precursor cells that can transplant specific tissues are also critical determinants in the effectiveness of HAC-based tissue replacement therapies. Translating the HAC-based gene therapy platform remains difficult, but ongoing animal model research will move the HAC platform closer to clinical trials.