Wang Chao, Sun Guanghong, Wang Ye, Kong Nana, Chi Yafei, Yang Leilei, Xin Qiliang, Teng Zhen, Wang Xu, Wen Yujun, Li Ying, Xia Guoliang
a State Key Laboratories for Agrobiotechnology and College of Biological Sciences, China Agricultural University , Beijing , China.
b School of Basic Medical Science, Capital Medical University , Beijing , China.
Drug Deliv. 2017 Nov;24(1):651-659. doi: 10.1080/10717544.2017.1293195.
Nano-scaled materials have been proved to be ideal DNA carriers for transgene. Bacterial magnetic particles (BMPs) help to reduce the toxicity of polyethylenimine (PEI), an efficient gene-transferring agent, and assist tissue transgene ex vivo. Here, the effectiveness of the BMP-PEI complex-conjugated foreign DNAs (BPDs) in promoting testes-mediated gene transfer (TMGT) in mouse was compared with that of liposome-conjugated foreign DNAs. The results proved that through testes injection, the clusters of BPDs successfully reached the cytoplasm and the nuclear of spermatogenesis cell, and expressed in testes of transgene founder mice. Additionally, the ratio of founder mice obtained from BPDs (88%) is about 3 times higher than the control (25%) (p < 0.05). Interestingly, the motility of sperms recovered from epididymis of the founder mice from BPD group were significantly improved, as compared with the control (p < 0.01). Based on classic breeding, the ratio of transgene mice within the first filial was significantly higher in BPDs compared with the control (73.8% versus 11.6%, p < 0.05). TMGT in this study did not produce visible histological changes in the testis. In conclusion, nano-scaled BPDs could be an alternative strategy for efficiently producing transgene mice in vivo.
纳米级材料已被证明是用于转基因的理想DNA载体。细菌磁性颗粒(BMPs)有助于降低高效基因转移剂聚乙烯亚胺(PEI)的毒性,并辅助离体组织转基因。在此,将BMP-PEI复合物偶联的外源DNA(BPDs)促进小鼠睾丸介导的基因转移(TMGT)的效果与脂质体偶联的外源DNA进行了比较。结果证明,通过睾丸注射,BPDs簇成功到达生精细胞的细胞质和细胞核,并在转基因奠基小鼠的睾丸中表达。此外,从BPDs获得的奠基小鼠比例(88%)约为对照组(25%)的3倍(p < 0.05)。有趣的是,与对照组相比,从BPD组奠基小鼠附睾中回收的精子活力显著提高(p < 0.01)。基于经典育种,BPDs组第一代子代中转基因小鼠的比例显著高于对照组(73.8%对11.6%,p < 0.05)。本研究中的TMGT未在睾丸中产生可见的组织学变化。总之,纳米级BPDs可能是在体内高效产生转基因小鼠的一种替代策略。