Bank A, Donovan-Peluso M, Lerner N, Rund D
Columbia University College of Physicians and Surgeons, Department of Medicine, New York, NY 10032.
Blood Cells. 1987;13(1-2):269-75.
Human globin genes can be transferred into mouse and human erythroid cells in culture, and can be appropriately expressed at the mRNA level in these cells. A plasmid containing a human beta globin gene is expressed in mouse erythroleukemia cells (MELC), and another containing a human epsilon or gamma gene is expressed in human erythroleukemia (K562) cells. A neomycin resistance (neoR) gene on the plasmids has been used to select for those cells containing the transferred globin genes; this selection may favor the expression of the globin genes by providing chromosomal positions requiring neoR expression. Analyzing clones resistant to G418, a neomycin analogue, demonstrated globin mRNA expression and induction. Retroviral vectors have also been used to transfer and appropriately express human beta genes in MELC. In addition, a plasmid containing a dihydrofolate reductase (DHFR) gene as well as neoR and beta globin genes has been used to amplify and express beta globin mRNA in MELC. These experiments suggest that high level appropriate expression of human beta globin genes is feasible and provides potentially useful approaches to the long-range goal of gene therapy for sickle cell anemia and beta thalassemia.
人类珠蛋白基因可在培养的小鼠和人类红细胞系细胞中进行转移，并能在这些细胞的mRNA水平上得到适当表达。含有人类β珠蛋白基因的质粒在小鼠红白血病细胞（MELC）中表达，而另一个含有人类ε或γ基因的质粒则在人类红白血病（K562）细胞中表达。质粒上的新霉素抗性（neoR）基因已被用于筛选那些含有转移珠蛋白基因的细胞；这种筛选可能通过提供需要neoR表达的染色体位置来促进珠蛋白基因的表达。分析对新霉素类似物G418有抗性的克隆，证实了珠蛋白mRNA的表达和诱导。逆转录病毒载体也已被用于在MELC中转移和适当表达人类β基因。此外，一个含有二氢叶酸还原酶（DHFR）基因以及neoR和β珠蛋白基因的质粒已被用于在MELC中扩增和表达β珠蛋白mRNA。这些实验表明，人类β珠蛋白基因的高水平适当表达是可行的，并为镰状细胞贫血和β地中海贫血基因治疗的长期目标提供了潜在有用的方法。
