Generation of pathogenic TPP1 mutations in human stem cells as a model for neuronal ceroid lipofuscinosis type 2 disease.
Generation of pathogenic TPP1 mutations in human stem cells as a model for neuronal ceroid lipofuscinosis type 2 disease.
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DOI：
10.1016/j.scr.2021.102323
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发表时间：
2021-05
影响因子：
1.2
通讯作者：
Morrow, M. Eric
中科院分区：
文献类型：
--
作者：
Ma, Li;Prada, M. Adriana;Schmidt, Michael;Morrow, M. Eric
神经元型蜡样脂褐素沉积症(CLN2病)是一种常染色体隐性遗传性神经退行性疾病，起病于2~4岁，以癫痫发作、视力丧失、进行性运动和智力衰退以及过早死亡为特征。CLN2病是由三肽基肽酶1(TPP1)基因功能丧失突变导致的TPP1酶活性缺陷引起的。大约60%的患者有两种致病变异之一(c.509-1G>；C或c.622C>；T[p.Arg208*])。为了建立CLN2病的人类干细胞模型，我们使用CRISPR/Cas9介导的敲入技术将这些纯合子状态的突变引入H9人类胚胎干细胞。还产生了c.622C>；T(p.(Arg208*))突变的杂合系，其中包括一个带有野生型等位基因的杂合突变体和由一个等位基因上的Indels产生的不同的复合杂合编码突变体。我们描述了导致这些线的产生的方法，并提供了关于这些CLN2疾病模型的初步验证和特征的数据。值得注意的是，处于纯合子状态的两个突变系(C.509-1G>；C和C.622C>；T[p.(Arg208*)])分别显示蛋白质减少或缺失，以及TPP1酶活性降低。这些模型，我们已经提供了广泛的共享，将有助于未来的分子和细胞机制的研究CLN2疾病和治疗开发。
Neuronal ceroid lipofuscinosis type 2 (CLN2 disease) is an autosomal recessive neurodegenerative disorder generally with onset at 2 to 4 years of age and characterized by seizures, loss of vision, progressive motor and mental decline, and premature death. CLN2 disease is caused by loss-of-function mutations in the tripeptidyl peptidase 1 (TPP1) gene leading to deficiency in TPP1 enzyme activity. Approximately 60% of patients have one of two pathogenic variants (c.509–1G > C or c.622C > T [p.(Arg208*)]). In order to generate a human stem cell model of CLN2 disease, we used CRISPR/Cas9-mediated knock-in technology to introduce these mutations in a homozygous state into H9 human embryonic stem cells. Heterozygous lines of the c.622C > T (p.(Arg208*)) mutation were also generated, which included a heterozygous mutant with a wild-type allele and different compound heterozygous coding mutants resulting from indels on one allele. We describe the methodology that led to the generation of the lines and provide data on the initial validation and characterization of these CLN2 disease models. Notably, both mutant lines (c.509–1G > C and c.622C > T [p.(Arg208*)]) in the homozygous state were shown to have reduced or absent protein, respectively, and deficiency of TPP1 enzyme activity. These models, which we have made available for wide-spread sharing, will be useful for future studies of molecular and cellular mechanisms underlying CLN2 disease and for therapeutic development.
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影响因子：
1.7
作者：
Kohlschuetter, Alfried;Schulz, Angela
通讯作者：
Schulz, Angela
H9细胞分化为神经外胚层的简单方法
影响因子：
2.6
作者：
Liu, Annuo;Zhang, Dijuan;Liu, Chao
通讯作者：
Liu, Chao
影响因子：
9.8
作者：
Sleat, DE;Gin, RM;Lobel, P
通讯作者：
Lobel, P
影响因子：
17.1
作者：
Lizarraga SB;Ma L;Maguire AM;van Dyck LI;Wu Q;Ouyang Q;Kavanaugh BC;Nagda D;Livi LL;Pescosolido MF;Schmidt M;Alabi S;Cowen MH;Brito-Vargas P;Hoffman-Kim D;Gamsiz Uzun ED;Schlessinger A;Jones RN;Morrow EM
通讯作者：
Morrow EM
DOI：
10.1073/pnas.90.2.532
发表时间：
1993-01-15
影响因子：
11.1
作者：
EPSTEIN, DJ;VOGAN, KJ;GROS, P
通讯作者：
GROS, P