Technology allowing efficient formation, delivery, isolation, and enrichment of single-copy Human Artificial Chromosome (HAC) for use in research, plant production, gene and cell therapy and more. Problem: Existing gene-editing techniques have limitations due to their reliance on viruses for delivery, restricting their packaging capacities. A more potent approach, HACs, can carry larger gene payloads alongside natural chromosomes without altering native genomes. However, a quarter century after the first human artificial chromosomes (HACs) were developed uncontrolled multimerization and DNA rearrangement still hinder their use. Solution: The inventors devised a method to produce HACs to create functional centromeres and engineered single-copy HACs allowing them to bypass the multimerization process. Furthermore, the inventors developed methodologies to deliver, isolate, and enrich these single-copy HACs, providing additional advantages such as stability, high rate of delivery, and high-efficiency production. This technology creates a potential for inserting multiple genes and gene cassettes with control elements into cells without altering the native genome. Potential Applications:
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Representative images of a single-copy HAC generated in HT1080 and U2OS cells. Intellectual Property:
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Docket # 23-10327