Wang Bin, Wang Jiale, Liu Yang, He Gordon, Jahan-Mihan Yasmin, Wang Yuxi, Hu Jie, Li Audrey, Muhammad Faheem, Bi Yan, Ji Baoan
Department of Cancer Biology, Mayo Clinic, Jacksonville, Florida, USA.
Department of Gastroenterology, Affiliated Drum Tower Hospital, Medical School of Nanjing University, Nanjing, China.
Genesis. 2025 Aug;63(4):e70022. doi: 10.1002/dvg.70022.
Tissue-specific gene manipulation using Cre/loxP or Flp/frt recombination systems is a cornerstone of genetically engineered mouse models. In this study, we aim to develop a novel hepatocyte-specific, tamoxifen-inducible Flp mouse line. BAC (bacterial artificial chromosome)-Alb(albumin)-FlpER2(estrogen receptor ligan binding domain) was developed by inserting IRES-FlpER2 cDNA between the translation stop codon and 3'-UTR of the mouse albumin gene in a bacterial artificial chromosome. Upon tamoxifen induction in mice crossed with reporter lines, western blotting, immunohistochemistry, immunofluorescence staining, and X-gal staining (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside Staining) were used to verify the recombination efficiency and specificity of this mouse model. Recombination was highly efficient and specific in hepatocytes, with no recombination detected in intrahepatic cholangiocytes or other organs in this mouse model. We generated a new tamoxifen-induced hepatocyte-specific mouse model with highly efficient recombination specifically in hepatocytes, and this model can be used to generate tumor model lines.
使用Cre/loxP或Flp/frt重组系统进行组织特异性基因操作是基因工程小鼠模型的基石。在本研究中，我们旨在开发一种新型的肝特异性、他莫昔芬诱导的Flp小鼠品系。通过将IRES-FlpER2 cDNA插入细菌人工染色体中鼠白蛋白基因的翻译终止密码子和3'-UTR之间，构建了BAC（细菌人工染色体）-Alb（白蛋白）-FlpER2（雌激素受体配体结合域）。在用他莫昔芬诱导与报告系杂交的小鼠后，使用蛋白质免疫印迹、免疫组织化学、免疫荧光染色和X-gal染色（5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷染色）来验证该小鼠模型的重组效率和特异性。在该小鼠模型中，重组在肝细胞中高效且特异，在肝内胆管细胞或其他器官中未检测到重组。我们生成了一种新的他莫昔芬诱导的肝特异性小鼠模型，其在肝细胞中具有高效的特异性重组，该模型可用于生成肿瘤模型系。
