Nagano Makoto, Watson Deborah J, Ryu Buom-Yong, Wolfe John H, Brinster Ralph L
Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104-6009, USA.
FEBS Lett. 2002 Jul 31;524(1-3):111-5. doi: 10.1016/s0014-5793(02)03010-7.
Transgenesis can be achieved in mice by retroviral transduction of male germ line stem cells (GSCs). However, the transduction efficiency by a Moloney murine leukemia virus (MMLV)-based vector is low, probably due to the characteristically slow cell cycle of stem cells. Since lentiviral vectors can transduce non-dividing cells, they have the potential to efficiently transduce GSCs. Here we report that male GSCs of mice can be transduced in vitro by a lentiviral vector and generate complete spermatogenesis when transplanted into infertile host testes. Transduction efficiencies were comparable to those for MMLV transduction using similar experimental conditions. The results suggest that both lentiviral and MMLV vectors could be effective in transducing GSCs of other species. In addition, these and previous studies suggest that transduction of immature donor stem cells transplanted into immature recipient testes will provide the most efficient system for male germ line modification.
通过逆转录病毒转导雄性生殖系干细胞(GSCs)可在小鼠中实现转基因。然而,基于莫洛尼鼠白血病病毒(MMLV)的载体的转导效率较低,这可能是由于干细胞具有特征性的缓慢细胞周期。由于慢病毒载体可转导非分裂细胞,它们有高效转导GSCs的潜力。在此我们报告,小鼠雄性GSCs可在体外被慢病毒载体转导,并在移植到不育宿主睾丸后产生完整的精子发生。在相似实验条件下,转导效率与MMLV转导相当。结果表明慢病毒和MMLV载体在转导其他物种的GSCs方面可能都是有效的。此外,这些研究以及之前的研究表明,将未成熟供体干细胞移植到未成熟受体睾丸中进行转导将为雄性生殖系修饰提供最有效的系统。