Department of Biomedicine, Aarhus University, Wilhelm Meyers Alle 4, DK-8000 Aarhus C, Denmark.
J Genet Genomics. 2012 Jun 20;39(6):269-74. doi: 10.1016/j.jgg.2012.05.004. Epub 2012 May 18.
Recombinant adeno-associated virus (rAAV) vectors have been extensively used for experimental gene therapy of inherited human diseases. Several advantages, such as simple vector construction, high targeting frequency by homologous recombination, and applicability to many cell types, make rAAV an attractive approach for targeted genome editing. Combined with cloning by somatic cell nuclear transfer (SCNT), this technology has recently been successfully adapted to generate gene-targeted pigs as models for cystic fibrosis, hereditary tyrosinemia type 1, and breast cancer. This review summarizes the development of rAAV for targeted genome editing in mammalian cells and provides strategies for enhancing the rAAV-mediated targeting frequency by homologous recombination. We discuss current development and application of the rAAV vectors for targeted genome editing in porcine primary fibroblasts, which are subsequently used as donor cells for SCNT to generate cloned genetically designed pigs and provide positive perspectives for the generation of gene-targeted pigs with rAAV in the future.
重组腺相关病毒 (rAAV) 载体已被广泛用于遗传性人类疾病的实验性基因治疗。rAAV 具有许多优点，例如载体构建简单、同源重组靶向频率高以及适用于多种细胞类型，使其成为靶向基因组编辑的一种有吸引力的方法。与体细胞核移植 (SCNT) 克隆相结合，该技术最近已成功应用于生成囊性纤维化、遗传性酪氨酸血症 1 型和乳腺癌的基因靶向猪模型。本综述总结了 rAAV 在哺乳动物细胞中靶向基因组编辑的发展，并提供了通过同源重组增强 rAAV 介导的靶向频率的策略。我们讨论了 rAAV 载体在猪原代成纤维细胞中的靶向基因组编辑的最新发展和应用，这些细胞随后被用作 SCNT 的供体细胞，以生成基因设计的克隆猪，并为未来使用 rAAV 生成基因靶向猪提供了积极的前景。
