Generation of IL17RB Knockout Cell Lines Using CRISPR/Cas9-Based Genome Editing.
Generation of IL17RB Knockout Cell Lines Using CRISPR/Cas9-Based Genome Editing.
复制标题
使用基于 CRISPR/Cas9 的基因组编辑生成 IL17RB 敲除细胞系。
DOI：
10.1007/978-1-0716-0247-8_28
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Zhu,Yan
中科院分区：
文献类型：
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作者：
Hu,Olivia;Provvido,Alessandro;Zhu,Yan
基于CRISPR/Cas9的基因组编辑是一种廉价而有效的遗传修饰工具。在这里，我们提出了一种使用CRISPR/Cas9介导的基因组缺失建立白细胞介素-17受体B（IL 17 RB）敲除细胞系的方法学方法。IL 17 RB基因编码特异性结合IL 17 B和IL 17 E的细胞因子受体，并在各种癌症中过表达。该方法涉及CRISPR设计、CRISPR克隆、将CRISPR克隆递送到细胞中，以及通过缺失筛选引物设计、基因组DNA提取和聚合酶链反应（PCR）来验证IL 17 RB基因缺失。类似的方法可用于产生具有其它感兴趣基因的基因敲除的哺乳动物细胞系。
CRISPR/Cas9-based genome editing is an inexpensive and efficient tool for genetic modification. Here we present a methodological approach of establishing interleukin-17 receptor B (IL17RB) knockout cell lines using CRISPR/Cas9-mediated genomic deletion. IL17RB gene encodes for a cytokine receptor that specifically binds to IL17B and IL17E and overexpressed in various cancers. The method involves CRISPR design, CRISPR cloning, delivery of CRISPR clone into cells, and verification of IL17RB gene deletion by deletion screening primer design, genomic DNA extraction, and polymerase chain reaction (PCR). Similar approaches can be used for generating mammalian cell lines with gene knockout for other genes of interest.