Advances in synthetic biology allow the design and manipulation of DNA from the scale of genes to genomes, enabling the engineering of complex genetic information for application in biomanufacturing, biomedicine and other areas. The transfer and subsequent maintenance of large DNA are two core steps in large scale genome rewriting. Compared to small DNA, the high molecular weight and fragility of large DNA make its transfer and maintenance a challenging process. This review outlines the methods currently available for transferring and maintaining large DNA in bacteria, fungi, and mammalian cells. It highlights their mechanisms, capabilities and applications. The transfer methods are categorized into general methods (, electroporation, conjugative transfer, induced cell fusion-mediated transfer, and chemical transformation) and specialized methods (, natural transformation, mating-based transfer, virus-mediated transfection) based on their applicability to recipient cells. The maintenance methods are classified into genomic integration (, CRISPR/Cas-assisted insertion) and episomal maintenance (, artificial chromosomes). Additionally, this review identifies the major technological advantages and disadvantages of each method and discusses the development for large DNA transfer and maintenance technologies.

中文翻译：

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细菌、真菌和哺乳动物细胞中大DNA的转移和维持的进展


合成生物学的进步使得DNA的设计和操作成为可能，从基因到基因组的规模，使得复杂的遗传信息工程能够应用于生物制造、生物医学和其他领域。大DNA的转移和随后的维护是大规模基因组重写的两个核心步骤。与小 DNA 相比，大 DNA 的高分子量和脆弱性使其转移和维护成为一个具有挑战性的过程。这篇综述概述了目前可用于在细菌、真菌和哺乳动物细胞中转移和维持大 DNA 的方法。它强调了它们的机制、功能和应用。根据对受体的适用性，转移方法分为通用方法（电穿孔、接合转移、诱导细胞融合介导的转移和化学转化）和专用方法（自然转化、交配转移、病毒介导的转染）细胞。维持方法分为基因组整合（CRISPR/Cas辅助插入）和附加型维持（人工染色体）。此外，本综述还确定了每种方法的主要技术优点和缺点，并讨论了大型 DNA 转移和维护技术的发展。