Mysliwiec Matthew R, Chen Junqin, Powers Patricia A, Bartley Christopher R, Schneider Michael D, Lee Youngsook
Department of Anatomy, School of Medicine and Public Health, University of Wisconsin, Madison, Wisconsin 53706, USA.
Genesis. 2006 Sep;44(9):407-11. doi: 10.1002/dvg.20221.
The jumonji (jmj) gene plays important roles in multiple organ development in mouse, including cardiovascular development. Since JMJ is expressed widely during mouse development, it is essential that conditional knockout approaches be employed to ablate JMJ in a tissue-specific manner to identify the cell lineage specific roles of JMJ. In this report, we describe the establishment of a jmj conditional null allele in mice by generating a loxP-flanked (floxed) jmj allele, which allows the in vivo ablation of jmj via Cre recombinase-mediated deletion. Gene targeting was used to introduce loxP sites flanking exon 3 of the jmj allele to mouse embryonic stem cells. Our results indicate that the jmj floxed allele converts to a null allele in a heart-specific manner when embryos homozygous for the floxed jmj allele and carrying the alpha-myosin heavy chain promoter-Cre transgene were analyzed by Southern and Northern blot analyses. Therefore, this mouse line harboring the conditional jmj null allele will provide a valuable tool for deciphering the tissue and cell lineage specific roles of JMJ.
Jumonji(jmj)基因在小鼠的多个器官发育中发挥重要作用,包括心血管发育。由于JMJ在小鼠发育过程中广泛表达,因此必须采用条件性基因敲除方法以组织特异性方式消除JMJ,从而确定JMJ在细胞谱系中的特定作用。在本报告中,我们描述了通过产生一个loxP侧翼(floxed)的jmj等位基因在小鼠中建立jmj条件性无效等位基因的过程,这使得能够通过Cre重组酶介导的缺失在体内消除jmj。利用基因打靶技术将loxP位点引入到小鼠胚胎干细胞中jmj等位基因的外显子3两侧。我们的结果表明,当通过Southern和Northern印迹分析对纯合携带floxed jmj等位基因并带有α-肌球蛋白重链启动子-Cre转基因的胚胎进行分析时,floxed的jmj等位基因会以心脏特异性方式转化为无效等位基因。因此,这种携带条件性jmj无效等位基因的小鼠品系将为解读JMJ在组织和细胞谱系中的特定作用提供有价值的工具。