CRISPR/Cas9 及其他序列特异性核酸酶产生的靶标和脱靶突变的检测。
Detection of on-target and off-target mutations generated by CRISPR/Cas9 and other sequence-specific nucleases.
机构信息
Institute for Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany.
Institute for Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany; Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstraße 6, 52074 Aachen, Germany.
出版信息
Biotechnol Adv. 2017 Jan-Feb;35(1):95-104. doi: 10.1016/j.biotechadv.2016.12.003. Epub 2016 Dec 21.
The development of customizable sequence-specific nucleases such as TALENs, ZFNs and the powerful CRISPR/Cas9 system has revolutionized the field of genome editing. The CRISPR/Cas9 system is particularly versatile and has been applied in numerous species representing all branches of life. Regardless of the target organism, all researchers using sequence-specific nucleases face similar challenges: confirmation of the desired on-target mutation and the detection of off-target events. Here, we evaluate the most widely-used methods for the detection of on-target and off-target mutations in terms of workflow, sensitivity, strengths and weaknesses.
定制化序列特异性核酸酶（如 TALEN、ZFN 以及强大的 CRISPR/Cas9 系统）的发展彻底改变了基因组编辑领域。CRISPR/Cas9 系统特别灵活，已应用于代表生命各个分支的众多物种中。无论目标生物是什么，所有使用序列特异性核酸酶的研究人员都面临着类似的挑战：确认所需的靶基因突变和检测脱靶事件。在这里，我们根据工作流程、灵敏度、优势和劣势，评估了最广泛使用的用于检测靶标和脱靶突变的方法。
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