Wakker Vincent, Brons Janynke F, Aanhaanen Wim T J, van Roon Marian A, Moorman Antoon F M, Christoffels Vincent M
Heart Failure Research Center, Academic Medical Center, Amsterdam, The Netherlands.
Genesis. 2010 Mar;48(3):195-9. doi: 10.1002/dvg.20596.
The T-box transcription factor Tbx2 plays important roles in patterning and development, and has been implicated in cell-cycle regulation and cancer. Conventional disruption of Tbx2 results in abnormalities of the heart, limbs, eye and other structures, and early fetal lethality. To gain insight into the role of Tbx2 in different tissues and at different stages of development, we have generated a conditional null allele of Tbx2 by flanking Exon 2 with loxP sites (Tbx2(fl2)). Homozygous Tbx2(fl2) mice are viable and fertile, indicating that the Tbx2(fl2) allele is a fully functional Tbx2 allele. Cre-mediated recombination, using a ubiquitously active CMV-Cre line, results in deletion of Exon 2 and loss of protein expression. Embryos homozygous for the recombined allele (Tbx2(Delta2)) show the same heart and limb defects as conventional Tbx2-deficient embryos. This Tbx2 conditional null allele will be a valuable tool to uncover tissue-specific roles of Tbx2 in development and disease.
T盒转录因子Tbx2在模式形成和发育过程中发挥着重要作用,并且与细胞周期调控及癌症有关。传统的Tbx2基因敲除会导致心脏、四肢、眼睛及其他结构出现异常,并导致早期胚胎致死。为深入了解Tbx2在不同组织及发育不同阶段的作用,我们通过在第2外显子两侧插入loxP位点构建了Tbx2的条件性无效等位基因(Tbx2(fl2))。纯合的Tbx2(fl2)小鼠可存活且可育,这表明Tbx2(fl2)等位基因是一个功能完全正常的Tbx2等位基因。利用广泛表达的CMV-Cre品系进行Cre介导的重组,会导致第2外显子缺失并使蛋白质表达丧失。重组等位基因(Tbx2(Delta2))的纯合胚胎表现出与传统Tbx2缺陷胚胎相同的心脏和四肢缺陷。这个Tbx2条件性无效等位基因将成为揭示Tbx2在发育和疾病中组织特异性作用的宝贵工具。