Mahata Barun, Biswas Kaushik
P1/12 CIT Scheme VIIM, Division of Molecular Medicine, Bose Institute, Kolkata, 700054, India.
Methods Mol Biol. 2017;1498:107-120. doi: 10.1007/978-1-4939-6472-7_7.
Precise and targeted genome editing using Transcription Activator-Like Effector Endonucleases (TALENs) has been widely used and proven to be an extremely effective and specific knockout strategy in both cultured cells and animal models. The current chapter describes a protocol for the construction and generation of TALENs using serial and hierarchical digestion and ligation steps, and using the synthesized TALEN pairs to achieve locus-specific targeted gene editing in mammalian cell lines using a modified clonal selection strategy in an easy and cost-efficient manner.
使用转录激活样效应物核酸酶（TALENs）进行精确且有针对性的基因组编辑已被广泛应用，并被证明在培养细胞和动物模型中都是一种极其有效且特异的基因敲除策略。本章描述了一种通过连续和分级消化及连接步骤构建和生成TALENs的方案，并使用合成的TALEN对，以一种简便且经济高效的方式，利用改良的克隆选择策略在哺乳动物细胞系中实现位点特异性靶向基因编辑。
