Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.
Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.
Nat Methods. 2023 Jul;20(7):1070-1081. doi: 10.1038/s41592-023-01896-x. Epub 2023 Jun 8.
The development of transgenic mouse models that express genes of interest in specific cell types has transformed our understanding of basic biology and disease. However, generating these models is time- and resource-intensive. Here we describe a model system, SELective Expression and Controlled Transduction In Vivo (SELECTIV), that enables efficient and specific expression of transgenes by coupling adeno-associated virus (AAV) vectors with Cre-inducible overexpression of the multi-serotype AAV receptor, AAVR. We demonstrate that transgenic AAVR overexpression greatly increases the efficiency of transduction of many diverse cell types, including muscle stem cells, which are normally refractory to AAV transduction. Superior specificity is achieved by combining Cre-mediated AAVR overexpression with whole-body knockout of endogenous Aavr, which is demonstrated in heart cardiomyocytes, liver hepatocytes and cholinergic neurons. The enhanced efficacy and exquisite specificity of SELECTIV has broad utility in development of new mouse model systems and expands the use of AAV for gene delivery in vivo.
表达目的基因的转基因小鼠模型的发展改变了我们对基础生物学和疾病的理解。然而，生成这些模型需要耗费大量的时间和资源。在这里，我们描述了一种模型系统，即选择性表达和体内控制转导（SELECTIV），它通过将腺相关病毒（AAV）载体与 Cre 诱导的多血清型 AAV 受体 AAVR 的过表达相结合，实现了转基因的高效和特异性表达。我们证明，转基因 AAVR 的过表达大大提高了许多不同细胞类型的转导效率，包括肌肉干细胞，这些细胞通常对 AAV 转导有抗性。通过将 Cre 介导的 AAVR 过表达与内源性 Aavr 的全身敲除相结合，可以实现更好的特异性，这在心脏心肌细胞、肝脏肝细胞和胆碱能神经元中得到了证明。SELECTIV 的增强功效和高度特异性在新型小鼠模型系统的开发中具有广泛的应用，并扩展了 AAV 在体内基因传递中的应用。
