Fekete D M, Cepko C L
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
Mol Cell Biol. 1993 Apr;13(4):2604-13. doi: 10.1128/mcb.13.4.2604-2613.1993.
Replication-competent avian retroviruses, capable of transducing and expressing up to 2 kb of nonviral sequences, are now available to effect widespread gene transfer in chicken (chick) embryos (S. H. Hughes, J. J. Greenhouse, C. J. Petropoulos, and P. Sutrave, J. Virol. 61:3004-3012, 1987). We have constructed novel avian retroviral vectors that encode human placental alkaline phosphatase as a marker whose expression can be histochemically monitored. These vectors have been tested for expression by introducing them into the embryonic chick nervous system. They have revealed that the expression of retrovirally transduced genes can be spatially and temporally limited without the need for tissue-specific promoters. By varying the site and time of infection, targeted gene transfer can be confined to selected populations of neural cells over the course of several days, a time window that is sufficient for many key developmental processes. The capability of differentially infecting specific target populations may avoid confounding variables such as detrimental effects of a transduced gene on processes unrelated to the cells or tissue of interest. These vectors and methods thus should be useful in studies of the effect of transduced genes on the development of various organs and tissues during avian embryogenesis. In addition, the vectors will facilitate studies aimed at an understanding of viral infection and expression patterns.
能够转导和表达长达2 kb非病毒序列的具有复制能力的禽逆转录病毒,现已可用于在鸡胚中实现广泛的基因转移(S. H. 休斯、J. J. 格林豪斯、C. J. 彼得罗普洛斯和P. 苏特拉夫,《病毒学杂志》61:3004 - 3012,1987年)。我们构建了新型禽逆转录病毒载体,其编码人胎盘碱性磷酸酶作为一种标志物,其表达可通过组织化学方法进行监测。通过将这些载体引入鸡胚神经系统来测试其表达情况。结果显示,逆转录病毒转导基因的表达可以在空间和时间上受到限制,而无需组织特异性启动子。通过改变感染的部位和时间,在几天的时间内,靶向基因转移可以局限于特定的神经细胞群体,这一时间段对于许多关键的发育过程来说是足够的。能够差异性地感染特定靶细胞群体的能力可以避免一些混淆变量,比如转导基因对与目标细胞或组织无关的过程产生的有害影响。因此,这些载体和方法在研究转导基因对禽胚胎发育过程中各种器官和组织的影响方面应该是有用的。此外,这些载体将有助于旨在了解病毒感染和表达模式的研究。