CRE MOUSE MODELS TO STUDY AMELOGENESIS

研究釉质形成的 CRE 小鼠模型

基本信息

批准号：
10674714
负责人：
Michael Lansdell Paine
金额：
$ 41.25万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10674714
关键词：
Age Ameloblasts Amelogenesis Animal Model Animals Anions Cell Line Cells Connexin 43 Crossbreeding Cystic Fibrosis Transmembrane Conductance Regulator Dental Dental Enamel Deposition Development Disadvantaged Disease Enamel Formation Enamel Organ Enterobacteria phage P1 Cre recombinase Funding Future Gene Expression Genes Grant Investments Keratin Knock-in Knockout Mice Knowledge LoxP-flanked allele Lung Maryland Modeling Multiple Organ Failure Mus Mutant Strains Mice National Institute of Dental and Craniofacial Research Organ Organoids Pancreas Pathology Pathway interactions Phase Proteins Publishing Research Research Personnel Resources Role Salivary Glands Skin Technology Testing The Jackson Laboratory Tissues Tongue Tooth structure Trachea ameloblastin animal data bronchial epithelium conditional knockout genomic locus in vivo Model mRNA Expression mouse Cre recombinase mouse model mutant novel promoter spatiotemporal tool

Age Ameloblasts Amelogenesis Animal Model Animals Anions Cell Line Cells Connexin 43 Crossbreeding Cystic Fibrosis Transmembrane Conductance Regulator Dental Dental Enamel Deposition Development Disadvantaged Disease Enamel Formation Enamel Organ Enterobacteria phage P1 Cre recombinase Funding Future Gene Expression Genes Grant Investments Keratin Knock-in Knockout Mice Knowledge LoxP-flanked allele Lung Maryland Modeling Multiple Organ Failure Mus Mutant Strains Mice National Institute of Dental and Craniofacial Research Organ Organoids Pancreas Pathology Pathway interactions Phase Proteins Publishing Research Research Personnel Resources Role Salivary Glands Skin Technology Testing The Jackson Laboratory Tissues Tongue Tooth structure Trachea ameloblastin animal data bronchial epithelium conditional knockout genomic locus in vivo Model mRNA Expression mouse Cre recombinase mouse model mutant novel promoter spatiotemporal tool

展开

项目摘要

PROJECT SUMMARY / ABSTRACT In June of 2017 a group of dental researchers met at the NIDCR to discuss the future of enamel research and concluded, in a summary statement, that in the field of enamel research, a lack of models to study enamel formation and disease has hampered recent progress. The group concluded that more appropriate ameloblast- like cell lines, investment in organoid and chip technology and novel animal models could be used to advance the field. With respect to animal models, I believe enamel researchers suffer from not having enamel organ- specific Cre recombinase mutant mouse. For the past 2 decades enamel researchers have used the Krt14-Cre (keratin 14-Cre recombinase) mutant to study enamel-specific activities by cross breeding with various loxP mouse lines. The significant disadvantage of using the Krt14-Cre mouse for enamel research is that Krt14 is expressed in multiple tissues including skin, bronchial epithelia, tongue, trachea, salivary glands, and many more organs, and because of this many of the developed loxP mouse lines are not appropriate to study amelogenesis. For example, mRNA expression levels of the anion exchanger protein (Slc4a2/AE2), or the cystic fibrosis transmembrane conductance regulator (Cftr), increases ~ 6-fold and 3-fold respectively in the enamel organ during maturation stage (compared to secretory stage), and beyond tooth formation both genes are widely expressed in lung and pancreas and are critical to their development. Both Slc4a2-null and Cftr-null mice have severe enamel pathologies. To use the Krt14-Cre mutant mouse to study the role of either AE2fl/fl or Cftrfl/fl would have significant limitations because these animals would predictably suffer from multiple organ failures at a young age. I propose to develop two animal models with a knockin of Cre recombinase into the ameloblastin (Ambn) and odontogenic ameloblast-associated (Odam) gene loci such that Cre expression is limited to secretory ameloblasts and maturation ameloblasts respectively. The UG3 stage of this grant will be devoted to the development of these two animal models; Ambn-Cre and Odam-Cre, and the UH3 phase will validate these two animals as unique in vivo models to study amelogenesis. At the completion of this project data from these animals will be published, and both lines deposited in an appropriate facility such as the Mutamt Mouse Resource and Research Centers (MMRRC).

项目摘要 /摘要 2017年6月，一群牙科研究人员在NIDCR开会，讨论搪瓷研究的未来和总结在摘要声明中总结说，在搪瓷研究领域，缺乏研究搪瓷的模型形成和疾病阻碍了最近的进步。该小组得出的结论是，更合适的ameleloblast- 像细胞系一样，可以使用对器官和芯片技术的投资以及新颖的动物模型来推进领域。关于动物模型，我相信搪瓷研究人员没有搪瓷器官特定的CRE重组酶突变小鼠。在过去的20年中，搪瓷研究人员使用了KRT14-CRE （角蛋白14-cre重组酶）突变体通过与各种LOXP交叉育种来研究牙釉质特异性活性鼠标线。使用KRT14-CRE小鼠进行搪瓷研究的重要缺点是Krt14是在多种组织中表达，包括皮肤，支气管上皮，舌，气管，唾液腺和许多更多的器官，因此，许多发达的LOXP小鼠系都不适合研究醒目。例如，阴离子交换器蛋白的mRNA表达水平（SLC4A2/AE2）或囊性纤维化跨膜电导调节剂（CFTR）分别增加约6倍和3倍成熟阶段（与分泌阶段相比）和牙齿形成超出两个基因的搪瓷器官在肺和胰腺中广泛表达，对它们的发展至关重要。 SLC4A2-NULL和CFTR-NULL都小鼠患有严重的搪瓷病理。使用KRT14-CRE突变小鼠研究AE2FL/FL或 CFTRFL/FL将有重大局限性，因为这些动物可以预见会遭受多个器官年轻时失败。我建议开发两个用CRE重组酶敲打的动物模型氨成蛋白（AMBN）和牙源性木质细胞相关（odam）基因基因座，使CRE表达为仅限于分泌的成成布和成熟的成熟成成木细胞。这笔赠款的UG3阶段将是致力于发展这两个动物模型； AMBN-CRE和ODAM-CRE，UH3阶段将验证这两只动物是研究障碍发生的独特体内模型。该项目完成时这些动物的数据将发布，两条线都沉积在适当的设施中，例如 MUTAMT小鼠资源和研究中心（MMRRC）。