Experimental studies on DNA transposable elements (TEs) have been limited in scale, leading to a lack of understanding of the factors influencing transposition activity, evolutionary dynamics, and application potential as genome engineering tools. We predicted 130 active DNA TEs from 102 metazoan genomes and evaluated their activity in human cells. We identified 40 active (integration-competent) TEs, surpassing the cumulative number (20) of TEs found previously. With this unified comparative data, we found that the Tc1/mariner superfamily exhibits elevated activity, potentially explaining their pervasive horizontal transfers. Further functional characterization of TEs revealed additional divergence in features such as insertion bias. Remarkably, in CAR-T therapy for hematological and solid tumors, Mariner2_AG (MAG), the most active DNA TE identified, largely outperformed two widely used vectors, the lentiviral vector and the TE-based vector SB100X. Overall, this study highlights the varied transposition features and evolutionary dynamics of DNA TEs and increases the TE toolbox diversity.

DNA转座元件（TE）的实验研究规模有限，导致人们对影响转座活性的因素、进化动力学以及作为基因组工程工具的应用潜力缺乏了解。我们预测了来自 102 个后生动物基因组的 130 个活性 DNA TE，并评估了它们在人类细胞中的活性。我们确定了 40 个活跃（有集成能力）的 TE，超过了之前发现的 TE 的累计数量（20 个）。通过这些统一的比较数据，我们发现 Tc1/水手超家族表现出较高的活性，这可能解释了它们普遍的水平转移。 TE 的进一步功能表征揭示了插入偏差等特征的额外差异。值得注意的是，在血液肿瘤和实体瘤的 CAR-T 疗法中，TE 鉴定出的最活跃的 DNA Mariner2_AG (MAG) 的性能很大程度上优于两种广泛使用的载体，即慢病毒载体和基于 TE 的载体 SB100X。总体而言，本研究强调了 DNA TE 的不同转座特征和进化动力学，并增加了 TE 工具箱的多样性。