Using CRISPR/Cas9 Technology for Manipulating Cell Death Regulators
Using CRISPR/Cas9 Technology for Manipulating Cell Death Regulators
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DOI：
10.1007/978-1-4939-3581-9_18
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发表时间：
2016-01-01
期刊：
影响因子：
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通讯作者：
Herold, Marco J.
中科院分区：
文献类型：
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作者：
Kueh, Andrew J.;Herold, Marco J.
有序的、规则间隔的短回文重复序列（CRISPR）/Cas9技术已被证明是在细胞系和小鼠中产生靶向突变的有用工具。然而，由于必需基因的突变，以组成型表达方式使用CRISPR/Cas9通常会导致低靶向效率和致死率。在这里，我们描述了诱导型慢病毒载体平台的使用，使得CRISPR/Cas9阳性细胞的快速转导和富集以及诱导后的高水平靶向突变成为可能。
Clustered, regularly interspaced, short palindromic repeats (CRISPR)/Cas9 technology has been demonstrated to be a useful tool for generating targeted mutations in cell lines and mice. However, the use of CRISPR/Cas9 in a constitutively expressed manner can often result in low targeting efficiencies and lethality due to mutations in essential genes. Here, we describe the use of an inducible lentiviral vector platform, enabling rapid transduction and enrichment of CRISPR/Cas9 positive cells and high levels of targeted mutations upon induction.