利用CRISPR/Cas系统快速高效构建血友病乙小鼠模型

doi:10.16288/j.yczz.15-117

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HEREDITAS(Beijing) ›› 2015, Vol. 37 ›› Issue (11): 1143-1148.doi: 10.16288/j.yczz.15-117

• Research Articles • Previous Articles Next Articles

A quick and efficient method to generate hemophilia B mouse models by the CRISPR/Cas system

Qihan Wang¹,Cong Huai¹,Ruilin Sun²,Hua Zhuang²,Hongyan Chen¹,Jian Fei²,Daru Lu¹

1. State Key Laboratory of Genetic Engineering, School of Life Science, Fudan University, Shanghai 200433, China;
2. Shanghai Research Center for Model Organisms, Shanghai 201203, China

Received:2015-03-22 Revised:2015-06-29 Online:2015-11-20 Published:2015-08-31

Abstract

Abstract: Hemophilia B, or the Christmas disease, is a common human disease caused by coagulation factor Ⅸ (FⅨ) deficiency. It is an X-linked recessive hereditary disease. Here we obtained FⅨ-knockout mouse strains with phenotype of hemophilia B with the CRISPR/Cas system efficiently. We chose the 8th exon as the target locus, and co-injected codon-optimized Cas9 mRNA with sgRNA of FⅨ into C57BL/6 mice zygotes. We obtained 60 mice in total and genotyped them by high resolution melting (HRM) and sequencing. The results showed the mutation rate was 85.0% in total, and 79.5% and 95.2% in males and females, respectively. No off-targets were detected in the similar locus by HRM. We future measured the FⅨ activity of each mice. The FⅨ: C of mutant mice were significantly below the normal level and reduced to 6.82% of wild-type mice. The activity assay demonstrated that all the mutant mice were lack of FⅨ. In summary, we have generated hemophilia B model mice with extreme efficiency, using the RNA-guided Cas9 nuclease gene editing system.

Key words: hemophilia B, CRISPR/Cas system, genome editing, mouse model

Qihan Wang,Cong Huai,Ruilin Sun,Hua Zhuang,Hongyan Chen,Jian Fei,Daru Lu. A quick and efficient method to generate hemophilia B mouse models by the CRISPR/Cas system[J]. HEREDITAS(Beijing), 2015, 37(11): 1143-1148.

References

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A quick and efficient method to generate hemophilia B mouse models by the CRISPR/Cas system

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