Strair R K, Towle M, Smith B R
Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06510.
Nucleic Acids Res. 1990 Aug 25;18(16):4759-62. doi: 10.1093/nar/18.16.4759.
Recombinant retroviruses have been utilized as vectors for gene transfer in model systems of gene therapy. Since many of these model systems require the transplantation of genetically modified primary cells it is important to devise methods which will allow the rapid and efficient selection for transplantation of only the cells which are capable of expressing high levels of the transferred gene. This report describes the use of beta-galactosidase as such a selectable marker. Bone marrow progenitors are infected with a recombinant retrovirus encoding beta-galactosidase. Using a fluorescence assay for beta-galactosidase we demonstrate that it is possible to use cell sorting to enrich for cells which will form bone marrow colonies that express high levels of beta-galactosidase. This rapid and non-toxic selection of bone marrow cells may facilitate attempts to achieve gene therapy in a variety of model systems.
重组逆转录病毒已被用作基因治疗模型系统中的基因转移载体。由于许多这些模型系统需要移植经过基因改造的原代细胞,因此设计出能够快速、高效地选择仅移植那些能够高水平表达转移基因的细胞的方法非常重要。本报告描述了使用β-半乳糖苷酶作为这样一种选择标记。骨髓祖细胞用编码β-半乳糖苷酶的重组逆转录病毒进行感染。通过针对β-半乳糖苷酶的荧光测定,我们证明可以利用细胞分选来富集那些能够形成高水平表达β-半乳糖苷酶的骨髓集落的细胞。这种对骨髓细胞的快速且无毒的选择可能有助于在各种模型系统中实现基因治疗的尝试。