转录激活样效应因子核酸酶介导的诱变与基因组编辑。
TALEN-Mediated Mutagenesis and Genome Editing.
作者信息
Ma Alvin C H, Chen Yi, Blackburn Patrick R, Ekker Stephen C
机构信息
Department of Medicine, LKS Faculty of Medicine, University of Hong Kong, Pokfulam, Hong Kong.
Mayo Clinic, Rochester, MN, USA.
出版信息
Methods Mol Biol. 2016;1451:17-30. doi: 10.1007/978-1-4939-3771-4_2.
Abstract
Transcription activator-like effectors (TALEs) are important genomic tools with customizable DNA-binding motifs for locus-specific modifications. In particular, TALE nucleases or TALENs have been successfully used in the zebrafish model system to introduce targeted mutations via repair of double-stranded breaks (DSBs) either through nonhomologous end joining (NHEJ) or by homology-directed repair (HDR) and homology-independent repair in the presence of a donor template. Compared with other customizable nucleases, TALENs offer high binding specificity and fewer sequence constraints in targeting the genome, with comparable mutagenic activity. Here, we describe a detailed in silico design tool for zebrafish genome editing for TALENs and CRISPR/Cas9 custom restriction enzymes using Mojo Hand 2.0 software.
摘要
转录激活样效应因子（TALEs）是重要的基因组工具，具有可定制的DNA结合基序，用于位点特异性修饰。特别是，TALE核酸酶或TALENs已成功应用于斑马鱼模型系统，通过非同源末端连接（NHEJ）或同源定向修复（HDR）修复双链断裂（DSB），以及在存在供体模板的情况下进行同源独立修复，从而引入靶向突变。与其他可定制核酸酶相比，TALENs在靶向基因组时具有高结合特异性和较少的序列限制，且诱变活性相当。在此，我们描述了一种使用Mojo Hand 2.0软件对斑马鱼基因组编辑进行TALENs和CRISPR/Cas9定制限制酶的详细计算机辅助设计工具。
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