Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195, USA, Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, WA 98101, USA, Pregenen, Inc., Seattle, WA 98103, USA, Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA, Cellectis S.A., Paris, 75013, France, Division of Basic Sciences, Fred Hutch Cancer Research Center, Seattle, WA 98109, USA, Department of Biochemistry, University of Washington, Seattle, WA 98195, USA, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA and Department of Immunology, University of Washington, Seattle, WA 98195, USA.
Nucleic Acids Res. 2014 Feb;42(4):2591-601. doi: 10.1093/nar/gkt1224. Epub 2013 Nov 26.
Rare-cleaving endonucleases have emerged as important tools for making targeted genome modifications. While multiple platforms are now available to generate reagents for research applications, each existing platform has significant limitations in one or more of three key properties necessary for therapeutic application: efficiency of cleavage at the desired target site, specificity of cleavage (i.e. rate of cleavage at 'off-target' sites), and efficient/facile means for delivery to desired target cells. Here, we describe the development of a single-chain rare-cleaving nuclease architecture, which we designate 'megaTAL', in which the DNA binding region of a transcription activator-like (TAL) effector is used to 'address' a site-specific meganuclease adjacent to a single desired genomic target site. This architecture allows the generation of extremely active and hyper-specific compact nucleases that are compatible with all current viral and nonviral cell delivery methods.
稀有切割内切酶已成为靶向基因组修饰的重要工具。虽然现在有多种平台可用于研究应用的试剂生成，但每个现有的平台在三个关键特性中的一个或多个方面都有显著的局限性，这三个关键特性对于治疗应用是必要的：在所需靶位点的切割效率、切割的特异性（即在“脱靶”位点的切割率），以及有效/简便的方法将其递送至所需的靶细胞。在这里，我们描述了一种单链稀有切割内切酶结构的开发，我们将其命名为“megaTAL”，其中转录激活因子样（TAL）效应物的 DNA 结合区用于“寻址”紧邻单个所需基因组靶位点的位点特异性 meganuclease。这种结构允许生成非常活跃和超特异性的紧凑型核酸酶，与所有当前的病毒和非病毒细胞递送方法兼容。
Zotero 插件
