Format

Send to

Choose Destination
EMBO Mol Med. 2016 May 2;8(5):477-88. doi: 10.15252/emmm.201506039. Print 2016 May.

CRISPR/Cas9-mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse.

Author information

1
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
2
Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Hainan Reproductive Medical Center, the Affiliated Hospital of Hainan Medical University Hainan Medical University, Haikou, China Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, China mayl1990@foxmail.com myliu@bio.ecnu.edu.cn dlli@bio.ecnu.edu.cn.
3
Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Hainan Reproductive Medical Center, the Affiliated Hospital of Hainan Medical University Hainan Medical University, Haikou, China.
4
Fengxian Hospital affiliated to Southern Medical University, Shanghai, China.
5
Bioray Laboratories Inc., Shanghai, China.
6
Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
7
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China Department of Molecular and Cellular Medicine, The Institute of Biosciences and Technology, Texas A&M University Health Science Center, Houston, TX, USA mayl1990@foxmail.com myliu@bio.ecnu.edu.cn dlli@bio.ecnu.edu.cn.
8
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China mayl1990@foxmail.com myliu@bio.ecnu.edu.cn dlli@bio.ecnu.edu.cn.

Abstract

The X-linked genetic bleeding disorder caused by deficiency of coagulator factor IX, hemophilia B, is a disease ideally suited for gene therapy with genome editing technology. Here, we identify a family with hemophilia B carrying a novel mutation, Y371D, in the human F9 gene. The CRISPR/Cas9 system was used to generate distinct genetically modified mouse models and confirmed that the novel Y371D mutation resulted in a more severe hemophilia B phenotype than the previously identified Y371S mutation. To develop therapeutic strategies targeting this mutation, we subsequently compared naked DNA constructs versus adenoviral vectors to deliver Cas9 components targeting the F9 Y371D mutation in adult mice. After treatment, hemophilia B mice receiving naked DNA constructs exhibited correction of over 0.56% of F9 alleles in hepatocytes, which was sufficient to restore hemostasis. In contrast, the adenoviral delivery system resulted in a higher corrective efficiency but no therapeutic effects due to severe hepatic toxicity. Our studies suggest that CRISPR/Cas-mediated in situ genome editing could be a feasible therapeutic strategy for human hereditary diseases, although an efficient and clinically relevant delivery system is required for further clinical studies.

KEYWORDS:

gene therapy; genome editing; hemophilia B; hemostasis; monogenetic disease

PMID:
26964564
PMCID:
PMC5125832
DOI:
10.15252/emmm.201506039
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Wiley Icon for PubMed Central
Loading ...
Support Center