Macromolecular Crystallography Group, Structural Biology and Biocomputing Programme, Spanish National Cancer Research Centre (CNIO), Melchor Fdez Almagro, Madrid, Spain.
Crit Rev Biochem Mol Biol. 2012 May-Jun;47(3):207-21. doi: 10.3109/10409238.2011.652358. Epub 2012 Jan 28.
The engineering of protein-DNA interactions in different protein scaffolds may provide "toolkits" to modify the genome. Homing endonucleases are powerful tools for genome manipulation through homologous recombination, as these enzymes possess a very low frequency of DNA cleavage in eukaryotic genomes due to their high specificity. Therefore, the combination of a precise "cutter" with the presence of a natural or modified homologous DNA donor provides a potentially useful means to modify the genome. However, the basis of protein-DNA recognition must be understood to generate tailored enzymes that target the DNA at sites of interest. The engineering of homing endonucleases and alternative scaffolds, such as zinc fingers or transcription activator-like effector domains, has demonstrated the potential of these approaches to create new specific instruments to target genes for inactivation or repair. Customized homing endonucleases targeting selected human genes can excise or correct regions of genes implicated in monogenic diseases, thereby representing important tools for intervention in eukaryotic genomes.
在不同蛋白质支架中设计蛋白质-DNA 相互作用，可能为修饰基因组提供“工具包”。归巢内切核酸酶是通过同源重组进行基因组操作的有力工具，因为这些酶由于其高度特异性，在真核基因组中 DNA 切割的频率非常低。因此，精确的“切割器”与天然或修饰的同源 DNA 供体的存在相结合，为修饰基因组提供了一种潜在的有用手段。然而，为了生成针对感兴趣的 DNA 位点的靶向酶，必须了解蛋白质-DNA 识别的基础。归巢内切核酸酶和替代支架（如锌指或转录激活因子样效应结构域）的设计已经证明了这些方法具有潜在的能力，可以创建新的特异性工具来靶向基因失活或修复。针对选定人类基因的定制归巢内切核酸酶可以切除或纠正单基因疾病相关基因的区域，因此代表了干预真核基因组的重要工具。
