基于CRISPR-Cas9构建树突状细胞相关C型凝集素-1基因敲除小鼠模型

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Abstract Objective To investigate the effect of the deletion of the dendritic cell-associated C-type lectin-1(Dectin-1)gene on the tumor growth in mice.Methods CRISPR-Cas9 gene editing technology was used to construct the Dectin-1 gene knockout mouse model.sgRNA targeting the gene of interest was designed to achieve specific cutting of DNA double strand by Cas9 protein.The F0 generation mice were obtained from C57BL/6 surrogate mice,which were fertilized egg micro-injected with sgRNA and ssDNA purified in vitro and Cas9,and Dectin-1 deficient mice were obtained from F0 generation mice by gene identification and DNA sequencing.The tumor models of Wild-type and Dectin-1 knockout mice were further established to evaluate Dectin-1 on the antitumor effect of beta-glucan.Results The Dectin-1 gene of C57BL/6 wild-type mice was successfully knocked out using CRISPR-Cas9 gene editing technology.The deletion of partial nucleotides of Dectin-1 gene was confirmed by PCR technology and DNA sequencing,and the functional experiment showed that the tumor growth rate of Dectin-1 knockout mice was higher than that of wild-type mice,the difference was statistically significant(P<0.05).Conclusion The Dectin-1 gene knockout mouse model is successfully constructed,and the mouse tumor model further confirms the effect of Dectin-1 on the antitumor of beta-glucan,which can lay a foundation for the further study of the role of Dectin-1 in tumor immunotherapy.

Key words： CRISPR-Cas9 gene editing technology Dendritic cell-associated C-type lectin-1 Tumor immunotherapy Beta-glucan

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	CHEN Jie
	XIE Ye-wen
	PAN Jie
	DING Jun

Cite this article:

CHEN Jie,XIE Ye-wen,PAN Jie, et al. Construction of Dectin-1 gene knockout mouse models based on CRISPRCas9[J]. 中国当代医药, 2021, 28(32): 40-44.

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http://dangdaiyiyao.com/EN/ OR http://dangdaiyiyao.com/EN/Y2021/V28/I32/40

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