Zhu Zaobing, Lu Shengtao, Wang Hongchun, Wang Fan, Xu Wenting, Zhu Yulei, Xue Jing, Yang Litao
Joint International Research Laboratory of Metabolic and Developmental Sciences, Yazhou Bay Institute of Deepsea Sci-Tech, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, People's Republic of China.
ACS Appl Mater Interfaces. 2025 Jan 15;17(2):2716-2735. doi: 10.1021/acsami.4c14208. Epub 2025 Jan 6.
Understanding the integration of transgene DNA (T-DNA) in transgenic crops, animals, and clinical applications is paramount for ensuring the stability and expression of inserted genes, which directly influence desired traits and therapeutic outcomes. Analyzing T-DNA integration patterns is essential for identifying potential unintended effects and evaluating the safety and environmental implications of genetically modified organisms (GMOs). This knowledge is crucial for regulatory compliance and fostering public trust in biotechnology by demonstrating transparency in genetic modifications. This review highlights recent advancements in T-DNA integration analysis, specifically focusing on targeted DNA enrichment and sequencing strategies. We examine key technologies, such as polymerase chain reaction (PCR)-based methods, hybridization capture, RNA/DNA-guided endonuclease-mediated enrichment, and high-throughput resequencing, emphasizing their contributions to enhancing precision and efficiency in transgene integration analysis. We discuss the principles, applications, and recent developments in these techniques, underscoring their critical role in advancing biotechnological products. Additionally, we address the existing challenges and future directions in the field, offering a comprehensive overview of how innovative DNA-targeted enrichment and sequencing strategies are reshaping biotechnology and genomics.
了解转基因DNA（T-DNA）在转基因作物、动物及临床应用中的整合情况，对于确保插入基因的稳定性和表达至关重要，因为这直接影响所需性状和治疗效果。分析T-DNA整合模式对于识别潜在的非预期效应以及评估转基因生物（GMO）的安全性和环境影响至关重要。这些知识对于监管合规以及通过展示基因改造的透明度来增强公众对生物技术的信任至关重要。本综述重点介绍了T-DNA整合分析的最新进展，特别关注靶向DNA富集和测序策略。我们研究了关键技术，如基于聚合酶链反应（PCR）的方法、杂交捕获、RNA/DNA引导的核酸内切酶介导的富集以及高通量重测序，强调了它们在提高转基因整合分析的精度和效率方面的贡献。我们讨论了这些技术的原理、应用和最新进展，强调了它们在推进生物技术产品方面的关键作用。此外，我们还探讨了该领域目前存在的挑战和未来方向，全面概述了创新的DNA靶向富集和测序策略如何重塑生物技术和基因组学。
