Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA.
Institute for Advanced Co-Creation Studies, Osaka University, Osaka, 560-8531, Japan.
Cell Res. 2019 Oct;29(10):804-819. doi: 10.1038/s41422-019-0213-0. Epub 2019 Aug 23.
In vivo genome editing represents a powerful strategy for both understanding basic biology and treating inherited diseases. However, it remains a challenge to develop universal and efficient in vivo genome-editing tools for tissues that comprise diverse cell types in either a dividing or non-dividing state. Here, we describe a versatile in vivo gene knock-in methodology that enables the targeting of a broad range of mutations and cell types through the insertion of a minigene at an intron of the target gene locus using an intracellularly linearized single homology arm donor. As a proof-of-concept, we focused on a mouse model of premature-aging caused by a dominant point mutation, which is difficult to repair using existing in vivo genome-editing tools. Systemic treatment using our new method ameliorated aging-associated phenotypes and extended animal lifespan, thus highlighting the potential of this methodology for a broad range of in vivo genome-editing applications.
体内基因组编辑代表了一种强大的策略，既可以用于理解基础生物学，也可以用于治疗遗传性疾病。然而，开发用于包含不同类型细胞的组织的通用和高效的体内基因组编辑工具仍然是一个挑战，这些组织处于分裂或非分裂状态。在这里，我们描述了一种通用的体内基因敲入方法，该方法通过使用细胞内线性化的单同源臂供体将一个小基因插入靶基因座的内含子，从而能够靶向广泛的突变和细胞类型。作为概念验证，我们专注于一种由显性点突变引起的过早衰老的小鼠模型，该模型使用现有的体内基因组编辑工具难以修复。使用我们的新方法进行全身治疗改善了与衰老相关的表型并延长了动物的寿命，从而突出了该方法在广泛的体内基因组编辑应用中的潜力。
