Banares Steven, Zeh Karin, Krajewska Maryla, Kermer Pawel, Baribault Helene, Reed John C, Krajewski Stan
The Burnham Institute, La Jolla, California 92037, USA.
Genesis. 2005 May;42(1):6-16. doi: 10.1002/gene.20117.
Tissue-specific gene ablation is accomplished by combining conventional gene targeting approaches with site-specific recombinases such as the Cre/loxP system. Despite the use of a cardiac-specific rat myosin light chain II promoter, our transgenic line (CRE3) had little or no Cre expression in the heart; however, strong Cre activity was detected in the brain as early as gestation day E11.5. This was determined by several methods including crossing our mouse line with a lacZ indicator line (ROSA26). Transgenic Cre, in this mouse line, mediated DNA recombination of loxP-flanked genes selectively in neurons throughout the gray matter of the brain, cerebellum, spinal cord, as well as retina, dorsal, and sympathetic ganglia. Cre protein was also detected by immunohistochemistry exclusively in neurons, but not in other types of cells or tissues. Thus, our transgenic CRE3 mice provide pan-neuronal expression of CRE for carrying out conditional deletion of genes in neurons and their progenitors.
组织特异性基因敲除是通过将传统的基因靶向方法与位点特异性重组酶（如Cre/loxP系统）相结合来实现的。尽管使用了心脏特异性的大鼠肌球蛋白轻链II启动子，但我们的转基因品系（CRE3）在心脏中几乎没有或没有Cre表达；然而，早在胚胎第11.5天就检测到大脑中有强烈的Cre活性。这是通过几种方法确定的，包括将我们的小鼠品系与lacZ指示品系（ROSA26）杂交。在这个小鼠品系中，转基因Cre介导loxP侧翼基因在整个大脑灰质、小脑、脊髓以及视网膜、背根神经节和交感神经节中的神经元中选择性地进行DNA重组。通过免疫组织化学也仅在神经元中检测到Cre蛋白，而在其他类型的细胞或组织中未检测到。因此，我们的转基因CRE3小鼠提供了CRE的全神经元表达，用于在神经元及其祖细胞中进行基因的条件性缺失。
