Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN, USA.
BMC Bioinformatics. 2013 Jan 16;14:1. doi: 10.1186/1471-2105-14-1.
Recent studies of transcription activator-like (TAL) effector domains fused to nucleases (TALENs) demonstrate enormous potential for genome editing. Effective design of TALENs requires a combination of selecting appropriate genetic features, finding pairs of binding sites based on a consensus sequence, and, in some cases, identifying endogenous restriction sites for downstream molecular genetic applications.
We present the web-based program Mojo Hand for designing TAL and TALEN constructs for genome editing applications (http://www.talendesign.org). We describe the algorithm and its implementation. The features of Mojo Hand include (1) automatic download of genomic data from the National Center for Biotechnology Information, (2) analysis of any DNA sequence to reveal pairs of binding sites based on a user-defined template, (3) selection of restriction-enzyme recognition sites in the spacer between the TAL monomer binding sites including options for the selection of restriction enzyme suppliers, and (4) output files designed for subsequent TALEN construction using the Golden Gate assembly method.
Mojo Hand enables the rapid identification of TAL binding sites for use in TALEN design. The assembly of TALEN constructs, is also simplified by using the TAL-site prediction program in conjunction with a spreadsheet management aid of reagent concentrations and TALEN formulation. Mojo Hand enables scientists to more rapidly deploy TALENs for genome editing applications.
近期研究表明转录激活因子样效应物结构域融合到核酸酶(TALENs)在基因组编辑中具有巨大的潜力。TALENs 的有效设计需要结合选择合适的遗传特征、根据共识序列找到结合位点对、以及在某些情况下识别用于下游分子遗传应用的内源性限制位点。
我们提出了一种用于基因组编辑应用的 TAL 和 TALEN 构建体设计的基于网络的程序 Mojo Hand(http://www.talendesign.org)。我们描述了其算法和实现。Mojo Hand 的特点包括:(1)自动从国家生物技术信息中心下载基因组数据,(2)分析任何 DNA 序列,以根据用户定义的模板揭示结合位点对,(3)在 TAL 单体结合位点之间的间隔区中选择限制酶识别位点,包括选择限制酶供应商的选项,以及(4)设计用于后续使用 Golden Gate 组装方法构建 TALEN 的输出文件。
Mojo Hand 可快速识别 TAL 结合位点,用于 TALEN 设计。通过结合 TAL 结合位点预测程序和试剂浓度和 TALEN 配方的电子表格管理辅助工具,TALEN 构建体的组装也得到简化。Mojo Hand 使科学家能够更快地部署 TALEN 用于基因组编辑应用。