Lewandoski M, Wassarman K M, Martin G R
Department of Anatomy and Program in Developmental Biology, School of Medicine, University of California, San Francisco, California 94143-0452, USA.
Curr Biol. 1997 Feb 1;7(2):148-51. doi: 10.1016/s0960-9822(06)00059-5.
The site-specific DNA recombinase Cre is being used to develop a new generation of tools for controlling gene expression in mice [1]. Cre mediates the recombination of two directly repeated target (loxP) sites to a single loxP site, with concomitant excision of the DNA segment flanked by the loxP sites (the 'floxed' DNA). Such recombination can function to activate a gene by excising a floxed DNA segment that blocks expression because it either separates the regulatory and coding sequences of the gene [2] or interrupts the gene's open reading frame. Conversely, DNA excision can inactivate a gene if an essential fragment of the gene is floxed [3]. Gene activation or inactivation in vivo can be achieved by mating two different animals, one carrying a 'target gene' with appropriately placed loxP sites and one carrying a cre transgene. In most cases, the specificity of the system is dependent upon stringent regulation of cre expression. We describe here a mouse line in which cre expression is controlled by regulatory sequences from the mouse zona pellucida 3 (Zp3) gene, which is normally expressed exclusively in the growing oocyte prior to the completion of the first meiotic division [4]. We show that in target-bearing Zp3-cre mice, Cre-mediated recombination of the target gene apparently occurs in 100 % of oocytes. Moreover, Cre activity is not detected in the somatic tissues of most target-bearing Zp3-cre mice. Potential uses for this mouse line are discussed.
位点特异性DNA重组酶Cre正被用于开发新一代在小鼠中控制基因表达的工具[1]。Cre介导两个同向重复的靶标(loxP)位点重组为单个loxP位点,同时切除loxP位点侧翼的DNA片段(“floxed”DNA)。这种重组可通过切除因分离基因的调控序列和编码序列[2]或中断基因的开放阅读框而阻断表达的floxed DNA片段来激活基因。相反,如果基因的一个必需片段是floxed的,DNA切除可使基因失活[3]。体内基因激活或失活可通过将两种不同的动物进行交配来实现,一种携带带有适当放置的loxP位点的“靶基因”,另一种携带cre转基因。在大多数情况下,该系统的特异性取决于cre表达的严格调控。我们在此描述一种小鼠品系,其中cre表达由小鼠透明带3(Zp3)基因的调控序列控制,该基因通常仅在第一次减数分裂完成之前的生长卵母细胞中表达[4]。我们表明,在携带靶标的Zp3-cre小鼠中,靶基因的Cre介导的重组显然在100%的卵母细胞中发生。此外,在大多数携带靶标的Zp3-cre小鼠的体细胞组织中未检测到Cre活性。本文讨论了该小鼠品系的潜在用途。