Reddy S T, Stoker A W, Bissell M J
Division of Cell and Molecular Biology, Lawrence Berkeley Laboratory, CA 94720.
Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10505-9. doi: 10.1073/pnas.88.23.10505.
Retroviruses are valuable tools in studies of embryonic development, both as gene expression vectors and as cell lineage markers. In this study early chicken blastoderm cells are shown to be permissive for infection by Rous sarcoma virus and derivative replication-defective vectors, and, in contrast to previously published data, these cells will readily express viral genes. In cultured blastoderm cells, Rous sarcoma virus stably integrates and is transcribed efficiently, producing infectious virus particles. Using replication-defective vectors encoding the bacterial lacZ gene, we further show that blastoderms can be infected in culture and in ovo. In ovo, lacZ expression is seen within 24 hr of virus inoculation, and by 96 hr stably expressing clones of cells are observed in diverse tissues throughout the embryo, including epidermis, somites, and heart, as well as in extraembryonic membranes. Given the rapid onset of vector expression and the broad range of permissive cell types, it should be feasible to use Rous sarcoma virus-derived retroviruses as early lineage markers and expression vectors beginning at the blastoderm stage of avian embryogenesis.
逆转录病毒在胚胎发育研究中是有价值的工具，既作为基因表达载体，又作为细胞谱系标记。在本研究中，早期鸡胚盘细胞被证明对劳氏肉瘤病毒及其衍生的复制缺陷型载体的感染具有易感性，并且与先前发表的数据相反，这些细胞会很容易地表达病毒基因。在培养的胚盘细胞中，劳氏肉瘤病毒稳定整合并高效转录，产生有感染性的病毒颗粒。使用编码细菌lacZ基因的复制缺陷型载体，我们进一步证明胚盘在培养物中和在卵内均可被感染。在卵内，接种病毒后24小时内即可见到lacZ表达，到96小时时，在整个胚胎的各种组织中，包括表皮、体节、心脏以及胚外膜中，均可观察到稳定表达的细胞克隆。鉴于载体表达的快速起始以及允许感染的细胞类型范围广泛，从禽类胚胎发育的胚盘阶段开始，使用劳氏肉瘤病毒衍生的逆转录病毒作为早期谱系标记和表达载体应该是可行的。
