Targeting of the human F8 at the multicopy rDNA locus in Hemophilia a patient-derived iPSCs using TALENickases
Targeting of the human F8 at the multicopy rDNA locus in Hemophilia a patient-derived iPSCs using TALENickases
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使用 TALENickase 将人类 F8 靶向血友病患者来源的 iPSC 中的多拷贝 rDNA 基因座。
DOI：
10.1016/j.bbrc.2016.02.083
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发表时间：
2016-03-25
影响因子：
3.1
通讯作者：
Liang, Desheng
中科院分区：
文献类型：
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作者：
Pang, Jialun;Wu, Yong;Liang, Desheng
血友病A(HA)是一种由于缺乏凝血因子VIII(FVIII)而引起的单基因疾病。这种缺陷可能导致自发性关节出血或危及生命的出血，但直到最近还没有治愈HA的方法。在这项研究中，我们从重症HA患者中分离出诱导多能干细胞(IPSCs)，并利用转录激活物样效应尼克酶(TALENickase)将第VIII因子基因(F8)靶向于HA-IPSCs中的多拷贝核糖体DNA(RDNA)，旨在弥补FVIII蛋白的不足。结果表明，外源F8的一个以上拷贝可以整合到rDNA基因座上。重要的是，我们在靶向HA-IPSCs中检测到外源F8mRNA和FVIII蛋白。分化为内皮细胞后，仍可检测到外源性FVIII蛋白。因此，多拷贝rDNA基因座可以作为患者来源的ipSCs基因治疗的有效靶点。这一策略为HA和其他单基因疾病提供了一种基于IPSCs的新的治疗选择。(C)2016 Elsevier Inc.保留所有权利。
Hemophilia A (HA) is a monogenic disease due to lack of the clotting factor VIII (FVIII). This deficiency may lead to spontaneous joint hemorrhages or life-threatening bleeding but there is no cure for HA until very recently. In this study, we derived induced pluripotent stem cells (iPSCs) from patients with severe HA and used transcription activator-like effector nickases (TALENickases) to target the factor VIII gene (F8) at the multicopy ribosomal DNA (rDNA) locus in HA-iPSCs, aiming to rescue the shortage of FVIII protein. The results revealed that more than one copy of the exogenous F8 could be integrated into the rDNA locus. Importantly, we detected exogenous F8 mRNA and FVIII protein in targeted HA-iPSCs. After they were differentiated into endothelial cells (ECs), the exogenous FVIII protein was still detectable. Thus, it is showed that the multicopy rDNA locus could be utilized as an effective target site in patient derived iPSCs for gene therapy. This strategy provides a novel iPSCs-based therapeutic option for HA and other monogenic diseases. (C) 2016 Elsevier Inc. All rights reserved.