CRISPR/Cas9-mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse.
CRISPR/Cas9-mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse.
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CRISPR/Cas9介导的新型凝血因子IX基因突变的体细胞校正可改善小鼠血友病
DOI：
10.15252/emmm.201506039
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发表时间：
2016-05
影响因子：
11.1
通讯作者：
Li D
中科院分区：
文献类型：
--
作者：
Guan Y;Ma Y;Li Q;Sun Z;Ma L;Wu L;Wang L;Zeng L;Shao Y;Chen Y;Ma N;Lu W;Hu K;Han H;Yu Y;Huang Y;Liu M;Li D
由凝血因子IX缺乏引起的X连锁遗传性出血性疾病血友病B是一种非常适合使用基因组编辑技术进行基因治疗的疾病。在这里，我们确定了一个家庭与血友病B携带一种新的突变，Y371 D，在人类F9基因。CRISPR/Cas9系统用于产生不同的遗传修饰小鼠模型，并证实新的Y371 D突变导致比先前鉴定的Y371 S突变更严重的血友病B表型。为了开发靶向这种突变的治疗策略，我们随后比较了裸DNA构建体与腺病毒载体，以在成年小鼠中递送靶向F9 Y371 D突变的Cas9组分。治疗后，接受裸DNA构建体的血友病B小鼠表现出肝细胞中超过0.56%的F9等位基因的校正，这足以恢复止血。相比之下，腺病毒递送系统导致更高的矫正效率，但由于严重的肝毒性而没有治疗效果。我们的研究表明，CRISPR/Cas介导的原位基因组编辑可能是人类遗传性疾病的一种可行的治疗策略，尽管进一步的临床研究需要一种有效且临床相关的递送系统。
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.
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影响因子：
64.8
作者：
Barzel, A.;Paulk, N. K.;Shi, Y.;Huang, Y.;Chu, K.;Zhang, F.;Valdmanis, P. N.;Spector, L. P.;Porteus, M. H.;Gaensler, K. M.;Kay, M. A.
通讯作者：
Kay, M. A.
影响因子：
56.9
作者：
KAY, MA;ROTHENBERG, S;WOO, SLC
通讯作者：
WOO, SLC
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
64.5
作者：
Niu, Yuyu;Shen, Bin;Sha, Jiahao
通讯作者：
Sha, Jiahao
影响因子：
4.2
作者：
Muruve, DA;Barnes, MJ;Libermann, TA
通讯作者：
Libermann, TA