Kwok W W, Schuening F, Stead R B, Miller A D
Proc Natl Acad Sci U S A. 1986 Jun;83(12):4552-5. doi: 10.1073/pnas.83.12.4552.
Amphotropic retroviral vectors containing either a mutant dihydrofolate reductase gene (DHFR) or the bacterial neomycin phosphotransferase gene (neo) were used to infect canine hemopoietic cells. We report successful transfer and expression of the DHFR and neo genes in canine hemopoietic progenitor cells (colony-forming units, granulocyte/macrophage) as measured by the ability of the viruses to confer resistance to either methotrexate or the aminoglycoside G418, respectively. Transfer was achieved in the absence of helper virus by using retrovirus packaging cell lines. Successful transfer of these genes into canine hemopoietic progenitor cells in vitro indicates the feasibility of gene transfer into canine marrow for autologous reconstitution. Studies of transfer of new genetic information into a large, outbred animal such as the dog will provide a preclinical model for future gene therapy in humans.
含有突变二氢叶酸还原酶基因(DHFR)或细菌新霉素磷酸转移酶基因(neo)的双嗜性逆转录病毒载体被用于感染犬造血细胞。我们报告了DHFR和neo基因在犬造血祖细胞(集落形成单位,粒细胞/巨噬细胞)中的成功转移和表达,这分别通过病毒赋予对甲氨蝶呤或氨基糖苷类G418抗性的能力来衡量。通过使用逆转录病毒包装细胞系在无辅助病毒的情况下实现了转移。这些基因在体外成功转移到犬造血祖细胞中,表明将基因转移到犬骨髓进行自体重建的可行性。将新的遗传信息转移到诸如狗这样的大型远交动物中的研究将为未来人类基因治疗提供一个临床前模型。