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六个同基因克隆的H9人胚胎干细胞系中RB1启动子的双等位基因和单等位基因缺失

Biallelic and monoallelic deletion of the RB1 promoter in six isogenic clonal H9 hESC lines.

作者信息

Döpper Hannah, Horstmann Marius, Menges Julia, Bozet Morgane, Kanber Deniz, Steenpass Laura

机构信息

Institute of Human Genetics, University Hospital Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147 Essen, Germany.

Institute of Human Genetics, University Hospital Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147 Essen, Germany.

出版信息

Stem Cell Res. 2020 May;45:101779. doi: 10.1016/j.scr.2020.101779. Epub 2020 Mar 29.

Abstract

Retinoblastoma is a childhood tumor of the retina that is caused mostly by biallelic inactivation of the tumor suppressor gene RB1. To generate a research resource, we abrogated expression of RB1 in H9 hESCs by CRISPR/Cas9 induced deletion of the RB1 promoter, either on one or on both alleles. This enables studies on the role of RB1 loss during differentiation, for example in differentiation towards neural retina. The generation of three isogenic lines per deletion state enables validation of phenotypic results in independent clonal lines.

摘要

视网膜母细胞瘤是一种儿童期视网膜肿瘤,主要由肿瘤抑制基因RB1的双等位基因失活引起。为了生成一种研究资源,我们通过CRISPR/Cas9诱导删除RB1启动子,在一个或两个等位基因上消除了H9人胚胎干细胞中RB1的表达。这使得能够研究RB1缺失在分化过程中的作用,例如在向神经视网膜分化的过程中。每个缺失状态下三个同基因系的产生能够在独立的克隆系中验证表型结果。

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