Yang N S, Burkholder J, Roberts B, Martinell B, McCabe D
Department of Mammalian Genetics, Agracetus, Middleton, WI 53562.
Proc Natl Acad Sci U S A. 1990 Dec;87(24):9568-72. doi: 10.1073/pnas.87.24.9568.
Chimeric chloramphenicol acetyltransferase and beta-galactosidase marker genes were coated onto fine gold particles and used to bombard a variety of mammalian tissues and cells. Transient expression of the genes was obtained in liver, skin, and muscle tissues of rat and mouse bombarded in vivo. Similar results were obtained with freshly isolated ductal segments of rat and human mammary glands and primary cultures derived from these explants. Gene transfer and transient expression were also observed in eight human cell culture lines, including cells of epithelial, endothelial, fibroblast, and lymphocyte origin. Using CHO and MCF-7 cell cultures as models, we obtained stable gene transfer at frequencies of 1.7 x 10(-3) and 6 x 10(-4), respectively. The particle bombardment technology thus provides a useful means to transfer foreign genes into a variety of mammalian somatic cell systems. The method is applicable to tissues in vivo as well as to isolated cells in culture and has proven effective with all cell or tissue types tested thus far. This technology may therefore prove to be applicable in various aspects of gene therapy.
嵌合氯霉素乙酰转移酶和β-半乳糖苷酶标记基因被包被在微小的金颗粒上,并用于轰击多种哺乳动物组织和细胞。在体内轰击的大鼠和小鼠的肝脏、皮肤和肌肉组织中获得了这些基因的瞬时表达。在新鲜分离的大鼠和人类乳腺导管段以及从这些外植体衍生的原代培养物中也得到了类似结果。在8种人类细胞系中也观察到了基因转移和瞬时表达,包括上皮细胞、内皮细胞、成纤维细胞和淋巴细胞来源的细胞。以CHO和MCF-7细胞培养物为模型,我们分别以1.7×10⁻³和6×10⁻⁴的频率获得了稳定的基因转移。粒子轰击技术因此提供了一种将外源基因转移到多种哺乳动物体细胞系统的有用方法。该方法适用于体内组织以及培养中的分离细胞,并且到目前为止已证明对所有测试的细胞或组织类型均有效。因此,这项技术可能在基因治疗的各个方面都有应用价值。