基于CRISPR-Cas9定向编辑CCL17和CCL22基因的研究

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Abstract Objective To design CRISPR/Cas9-gRNA targeting human CCL17 and CCL22 gene,and to evaluate the editing effectiveness of the sgRNA.Methods The targeted editing sites of CCL17 and CCL22 genes were screened using tools provided by CRISPR website,then the knockout plasmid of CRISPR-Cas9-CCL17 and CRISPR-Cas9-CCL22 were constructed and separately transfected into 293 T cells.Finally,the T-A editing and sequencing methods of genomic PCR products were used to verify the editing efficiency of the designed sgRNA knockout plasmid.ResultsThe knockout plasmid of CRISPR-sgRNA targeting human CCL17 and CCL22 gene was successfully constructed with high editing efficiency,reached more than 70%.Conclusion The CRISPR-Cas9-CCL17 and CRISPR-Cas9-CCL22 show good editing efficiency,which provide foundation for the deep research of the function on CCL17 and CCL22.

Key words： Clustered regularly interspaced short palindromic reapeat sgRNA Chemokines Genome editing

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	GAO Shu-hui
	QU Chuang
	WU Feng-lin
	SHEN Han

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GAO Shu-hui,QU Chuang,WU Feng-lin, et al. Study on genome editing targeting CCL17 and CCL22 using CRISPRCas9 system[J]. 中国当代医药, 2018, 25(12): 4-8.

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http://dangdaiyiyao.com/EN/ OR http://dangdaiyiyao.com/EN/Y2018/V25/I12/4

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