Bowtell D D, Johnson G R, Kelso A, Cory S
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Mol Biol Med. 1987 Aug;4(4):229-50.
We have compared several recombinant retroviruses carrying the selective marker NeoR for gene delivery to murine haemopoietic stem cells. Provirus content and expression was assessed in animals reconstituted with infected bone marrow cells. With viruses bearing only a NeoR gene, viral RNA was readily detectable both in spleen colonies and in the haemopoietic tissues of long-term reconstituted mice. Thus, the progeny of infected primitive multipotential cells are competent to express integrated proviruses. The level of expression was significantly greater for proviruses carrying a modified long terminal repeat in which the Moloney enhancer had been replaced with that from the myeloproliferative sarcoma virus. With viruses bearing the c-myc gene in addition to NeoR, however, no viral RNA could be detected in spleen colonies harbouring proviral inserts, even though these viruses are expressed in vitro within haemopoietic cells. The implications of these results for gene transfer to haemopoietic stem cells are discussed.
我们比较了几种携带选择标记NeoR的重组逆转录病毒，用于将基因导入小鼠造血干细胞。在用感染的骨髓细胞重建的动物中评估了前病毒含量和表达情况。对于仅携带NeoR基因的病毒，在脾脏集落和长期重建小鼠的造血组织中都很容易检测到病毒RNA。因此，受感染的原始多能细胞的后代能够表达整合的前病毒。对于携带修饰的长末端重复序列的前病毒，其表达水平显著更高，其中莫洛尼氏增强子已被髓性增殖性肉瘤病毒的增强子所取代。然而，对于除NeoR外还携带c-myc基因的病毒，在含有前病毒插入片段的脾脏集落中未检测到病毒RNA，尽管这些病毒在体外造血细胞中表达。讨论了这些结果对造血干细胞基因转移的意义。
