Development of technology for efficient homologous recombination by the viral vector and zincfinger nuclease

近年来，基因组编辑技术如ZFN、TALEN、Cas9-CRISPR等的发展取得了很大进展，使我们能够在不需要大量劳动的情况下制备基因打靶小鼠，这些基因组编辑系统可以切割基因组上的特定序列，然后将DNA修复机制募集到消化位点，增强同源重组。另一方面，通过这些系统进行基因组编辑需要同时表达几种组分，这是这些系统在原代细胞中进行有效基因组编辑的缺点。在这项研究中，我们试图生成包含基因编辑系统多个组件的单个慢病毒载体，以实现高效的基因编辑。

Recent great progress of development of genome editing technology such as ZFN, TALEN, Cas9-CRISPR enabled us to generate gene targeting mice without labor intensive work.These genome editing systems can cut the specific sequence on the genome followed by recruitment of DNA repair machinery to the digested site and enhance the homologous recombination. On the other hand, simultaneous expression of several components are necessary for genome editing by these systems and this is the drawback of these systems for efficient genome editing in primary cells. In this study, we tried to generate single lentiviral vector harboring several components of gene editing system for efficient gene edition.