Applied StemCell, Inc., Milpitas, CA, USA.
Transgenic Res. 2021 Feb;30(1):91-104. doi: 10.1007/s11248-020-00226-7. Epub 2021 Jan 22.
The goal of this study is to establish a Cre/loxP rat resource for conditional and physiologically predictive rat models of human diseases. The laboratory rat (R. norvegicus) is a central experimental animal in several fields of biomedical research, such as cardiovascular diseases, aging, infectious diseases, autoimmunity, cancer models, transplantation biology, inflammation, cancer risk assessment, industrial toxicology, pharmacology, behavioral and addiction studies, and neurobiology. Up till recently, the ability of creating genetically modified rats has been limited compared to that in the mouse mainly due to lack of genetic manipulation tools and technologies in the rat. Recent advances in nucleases, such as CRISPR/Cas9 (clustered regularly-interspaced short palindromic repeats/CRISPR associated protein 9), as well as TARGATT™ integrase system enables fast, efficient and site-specific introduction of exogenous genetic elements into the rat genome. Here, we report the generation of a collection of tissue-specific, inducible transgenic Cre rats as tool models using TARGATT™, CRISPR/Cas9 and random transgenic approach. More specifically, we generated Cre driver rat models that allow controlled gene expression or knockout (conditional models) both temporally and spatially through the Cre-ERT2/loxP system. A total of 10 Cre rat lines and one Cre reporter/test line were generated, including eight (8) Cre lines for neural specific and two (2) lines for cardiovascular specific Cre expression. All of these lines have been deposited with the Rat Resource and Research Center and provide a much-needed resource for the bio-medical community who employ rat models for their studies of human diseases.
本研究旨在建立 Cre/loxP 大鼠资源,用于构建人类疾病的条件性和生理预测性大鼠模型。实验室大鼠(R. norvegicus)是心血管疾病、衰老、传染病、自身免疫、癌症模型、移植生物学、炎症、癌症风险评估、工业毒理学、药理学、行为和成瘾研究以及神经生物学等多个生物医学研究领域的重要实验动物。直到最近,由于缺乏大鼠遗传操作工具和技术,与小鼠相比,大鼠基因修饰的能力一直受到限制。近年来,核酸酶(如 CRISPR/Cas9(成簇规律间隔短回文重复/CRISPR 相关蛋白 9))和 TARGATT™整合酶系统的进步,使得快速、高效和定点地将外源遗传元件引入大鼠基因组成为可能。在这里,我们报告了使用 TARGATT™、CRISPR/Cas9 和随机转基因方法生成组织特异性、诱导型转基因 Cre 大鼠作为工具模型的集合。更具体地说,我们生成了 Cre 驱动大鼠模型,通过 Cre-ERT2/loxP 系统实现了时空可控的基因表达或敲除(条件性模型)。总共生成了 10 条 Cre 大鼠品系和 1 条 Cre 报告/测试品系,包括 8 条用于神经特异性和 2 条用于心血管特异性 Cre 表达的 Cre 大鼠品系。所有这些品系都已存入 Rat Resource and Research Center,为使用大鼠模型研究人类疾病的生物医学社区提供了急需的资源。