Establishment of liver⁃specific Lcat knock⁃in mouse models by the CRISPER/Cas9 technique
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Q785

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    Abstract:

    Objective:To construct lecithin - cholesterolacyl transferase(LCAT)knock -in mice by CRISPR/Cas9 - mediated gene editing and to obtain liver-specific overexpression of Lcat mice by mating with liver-specific Cre-expressing transgenic mice. Providing an animal model for the study of the mechanism of the Lcat gene in the development of liver-related metabolic diseases. Methods:Lcat knock -in mice were constructed by CRISPR/Cas9 technology;Liver - specific Cre- expressing transgenic mice were mated with Lcat knock -in mice to obtain liver -specific overexpressing Lcat mice;Genotyping mice by PCR;Quantitative real -time PCR(qPCR)and Western blot(WB)techniques were used to verify the expression of Lcat gene in C57BL/6 mice. Results:The PCR results showed that the liver-specific overexpression of Lcat gene in mice was successfully constructed;the qPCR results showed that the Lcat gene was specifically highly expressed in the liver,and the liver of knock -in mice showed higher Lcat expression;the WB results showed that LCAT protein was more highly expressed in the liver of liver-specific Lcat knock-in mice. Conclusion:Liver-specific overexpression of Lcat gene mice were successfully constructed,providing a platform for exploring the function of the Lcat gene at animal level in liver-related metabolic diseases and the associated pathogenesis.

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张芙蓉,苟黎明,李妍,王泽勇,杨斐,吴菁,覃健,薛斌.利用CRISPER/Cas9技术构建肝脏特异性敲入Lcat基因小鼠[J].南京医科大学学报(自然科学版英文版),2023,(10):1366-1371.

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  • Received:January 28,2023
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  • Online: October 23,2023
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