Lin Yongshun, Liu Guoqin, Wang Fen
Center for Cancer Biology and Nutrition, Institute of Biosciences and Technology, Texas A&M University System Health Science Center, Houston, Texas 77030-3303, USA.
Genesis. 2006 Mar;44(3):150-4. doi: 10.1002/gene.20194.
The fibroblast growth factor (FGF) family consists of 22 widely expressed regulatory polypeptides and controls a broad spectrum of cellular processes. Accumulating data show that FGF9 plays important roles both in embryogenesis and in adult tissue homeostasis. Ablation of Fgf9 alleles leads to lethality at the neonatal stage mainly due to malformations of the lung, as well as causing male-to-female sex reversal. To circumvent the neonatal lethality resulting from disruption of the Fgf9 gene, which hinders further characterization of the role of FGF9 in adult tissue function and homeostasis, we generated an Fgf9 conditional null allele for spatiotemporal- and tissue-specific disruption of Fgf9. Using gene targeting in mouse embryonic stem (ES) cells, we introduced two loxP sites flanking exon 1 in the Fgf9 allele, which encodes 93 amino acid residues at the N-terminal of FGF9. Our results indicate that the Fgf9 conditional null allele is a true conditional null that encodes wildtype activity and reverts to a null allele after recombination mediated by the Cre recombinase.
成纤维细胞生长因子(FGF)家族由22种广泛表达的调节性多肽组成,可控制广泛的细胞过程。越来越多的数据表明,FGF9在胚胎发育和成年组织稳态中均发挥重要作用。Fgf9等位基因的缺失会导致新生儿期死亡,主要原因是肺部畸形,同时还会导致雄性向雌性的性逆转。为了规避因Fgf9基因破坏导致的新生儿致死性(这阻碍了对FGF9在成年组织功能和稳态中作用的进一步研究),我们构建了一个Fgf9条件性无效等位基因,用于在时空和组织特异性水平上破坏Fgf9。利用小鼠胚胎干细胞(ES细胞)中的基因打靶技术,我们在Fgf9等位基因的外显子1两侧引入了两个loxP位点,该外显子编码FGF9 N端的93个氨基酸残基。我们的结果表明,Fgf9条件性无效等位基因是一个真正的条件性无效基因,它编码野生型活性,并在Cre重组酶介导的重组后恢复为无效等位基因。