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使用 CRISPR/Cas9 核糖核蛋白复合物对诱导多能干细胞进行基因组编辑,以模拟遗传性眼部疾病。

Genome Editing of Induced Pluripotent Stem Cells Using CRISPR/Cas9 Ribonucleoprotein Complexes to Model Genetic Ocular Diseases.

机构信息

The Ocular Genomics Institute at Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA.

出版信息

Methods Mol Biol. 2022;2549:321-334. doi: 10.1007/7651_2021_409.

Abstract

Genome editing with the use of CRISPR/Cas9 ribonucleoprotein complexes of induced pluripotent stem cells can be used to model many diseases. The combination of stem cells and gene editing technologies is a valuable tool to study ocular disorders, as many have been identified to be caused by specific genetic mutations. This protocol provides experimentally derived guidelines for genome editing of human induced pluripotent stem cells (iPSCs) using CRISPR/Cas9 ribonucleoprotein complexes to generate iPSCs harboring single nucleotide variants from ocular disorders. Edited iPSC can be further differentiated into retinal cells in order to study disease mechanisms as well as screen potential therapies.

摘要

利用 CRISPR/Cas9 核糖核蛋白复合物对诱导多能干细胞进行基因组编辑可用于模拟多种疾病。干细胞与基因编辑技术的结合是研究眼部疾病的一种有价值的工具,因为许多眼部疾病已被确定是由特定的基因突变引起的。本方案提供了使用 CRISPR/Cas9 核糖核蛋白复合物对人类诱导多能干细胞(iPSC)进行基因组编辑的实验衍生指南,以生成携带眼部疾病单核苷酸变异的 iPSC。编辑后的 iPSC 可进一步分化为视网膜细胞,以便研究疾病机制和筛选潜在的治疗方法。

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