Núcleo de Biotecnología, Centro de Desenvolvlmento Tecnológico, Campus Universitario, Unlversldade Federal de Pelotas, Pelotas, RS, Brazil.
Biol Res. 2011;44(3):229-34. Epub 2011 Nov 7.
Testis-mediated gene transfer (TMGT) has been used as in vivo gene transfer technology to introduce foreign DNA directly into testes, allowing mass gene transfer to offspring via mating. In this study, we used plasmid DNA (pEGFP-N1) mixed with dimethylsulfoxide (DMSO), N,N-dimethylacetamide (DMA) or liposome (Lipofectin) in an attempt to improve TMGT. Males receiving consecutive DNA complex injections were mated to normal females to obtain F0 progeny. In vivo evaluation of EGFP expression, RT-PCR and PCR were used to detect the expression and the presence of exogenous DNA in the progeny. We also evaluated possible testicular damage by histological procedures. PC R and RT-PCR analyses revealed that liposome and DMSO increased the rate of TMGT. Histological analyses demonstrated that repeated (4 times) injections of DNA complexes can affect spermatogenesis. DMSO was the most deleterious among the reagents tested. In this study, we detected the presence of transgene in the progeny, and its expression in blood cells. Consecutive injections of DNA complexes were associated with impaired spermatogenesis, suggesting requirement of optimal conditions for DNA delivery through TMGT.
睾丸介导的基因转移（TMGT）已被用作体内基因转移技术，将外源 DNA 直接导入睾丸，通过交配将大量基因转移给后代。在这项研究中，我们使用质粒 DNA（pEGFP-N1）与二甲基亚砜（DMSO）、N，N-二甲基乙酰胺（DMA）或脂质体（Lipofectin）混合，试图改进 TMGT。接受连续 DNA 复合物注射的雄性与正常雌性交配以获得 F0 后代。通过体内评估 EGFP 表达、RT-PCR 和 PCR 来检测后代中外源 DNA 的表达和存在。我们还通过组织学程序评估了可能的睾丸损伤。PCR 和 RT-PCR 分析表明脂质体和 DMSO 增加了 TMGT 的效率。组织学分析表明，重复（4 次）注射 DNA 复合物会影响精子发生。在所测试的试剂中，DMSO 是最具危害性的。在这项研究中，我们检测到了后代中转基因的存在及其在血细胞中的表达。连续注射 DNA 复合物与精子发生受损有关，这表明需要优化条件才能通过 TMGT 进行 DNA 传递。
