The N-terminal region of centrosomal protein 290 (CEP290) restores vision in a zebrafish model of human blindness.
The N-terminal region of centrosomal protein 290 (CEP290) restores vision in a zebrafish model of human blindness.
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DOI：
10.1093/hmg/ddr025
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发表时间：
2011-04-15
影响因子：
3.5
通讯作者：
Slusarski DC
中科院分区：
文献类型：
--
作者：
Baye LM;Patrinostro X;Swaminathan S;Beck JS;Zhang Y;Stone EM;Sheffield VC;Slusarski DC
编码中心体蛋白 290 (CEP290) 的基因是一种大型多结构域蛋白，是非综合征性致盲疾病莱伯先天性黑蒙 (LCA) 中最常见的突变基因。 CEP290 还与多种纤毛相关综合征有关，包括 Meckel-Gruber 综合征、Joubert 综合征、Senor-Loken 综合征和 Bardet-Biedl 综合征 (BBS)。在这项研究中，我们描述了 cep290 在斑马鱼中的发育和功能作用。反义寡核苷酸 [Morpholino (MO)] 旨在生成改变的 cep290 剪接产物，模拟最常见的 LCA 突变，用于基因敲低。我们发现，注射 cep290 MO 的胚胎减少了 Kupffer 囊泡的大小并延迟了黑素体运输，这是在斑马鱼中敲除 bbs 基因后观察到的两种表型。与纤毛功能中的作用一致，注射 cep290 MO 的胚胎表现出弯曲的体轴。由 CEP290 突变引起的 LCA 患者的视觉感知能力下降，尽管他们的视网膜完全分层。同样，对注射 cep290 MO 的斑马鱼的视网膜进行组织学检查，未发现明显的分层缺陷，但胚胎的视觉功能在统计学上显着下降。最后，我们证明，由 cep290 破坏引起的视力障碍可以通过仅表达人 CEP290 蛋白的 N 末端区域来挽救。这些数据表明，CEP290蛋白的特定区域足以恢复视觉功能，并且该区域可能是具有CEP290突变的LCA患者的可行基因治疗靶点。
The gene coding for centrosomal protein 290 (CEP290), a large multidomain protein, is the most frequently mutated gene underlying the non-syndromic blinding disorder Leber's congenital amaurosis (LCA). CEP290 has also been implicated in several cilia-related syndromic disorders including Meckel–Gruber syndrome, Joubert syndrome, Senor–Loken syndrome and Bardet–Biedl syndrome (BBS). In this study, we characterize the developmental and functional roles of cep290 in zebrafish. An antisense oligonucleotide [Morpholino (MO)], designed to generate an altered cep290 splice product that models the most common LCA mutation, was used for gene knockdown. We show that cep290 MO-injected embryos have reduced Kupffer's vesicle size and delays in melanosome transport, two phenotypes that are observed upon knockdown of bbs genes in zebrafish. Consistent with a role in cilia function, the cep290 MO-injected embryos exhibited a curved body axis. Patients with LCA caused by mutations in CEP290 have reduced visual perception, although they present with a fully laminated retina. Similarly, the histological examination of retinas from cep290 MO-injected zebrafish revealed no gross lamination defects, yet the embryos had a statistically significant reduction in visual function. Finally, we demonstrate that the vision impairment caused by the disruption of cep290 can be rescued by expressing only the N-terminal region of the human CEP290 protein. These data reveal that a specific region of the CEP290 protein is sufficient to restore visual function and this region may be a viable gene therapy target for LCA patients with mutations in CEP290.
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DOI：
10.1073/pnas.0600158103
发表时间：
2006-04-18
影响因子：
11.1
作者：
Chiang, AP;Beck, JS;Sheffield, VC
通讯作者：
Sheffield, VC
DOI：
10.1073/pnas.0500095102
发表时间：
2005-03-01
影响因子：
11.1
作者：
Deretic, D;Williams, AH;Arendt, A
通讯作者：
Arendt, A
影响因子：
6.1
作者：
O'Toole, John F.;Otto, Edgar A.;Hildebrandt, Friedhelm
通讯作者：
Hildebrandt, Friedhelm
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF
影响因子：
30.8
作者：
Otto, EA;Loeys, B;Hildebrandt, F
通讯作者：
Hildebrandt, F