Generation of human embryonic stem cell line with heterozygous RB1 deletion by CRIPSR/Cas9 nickase

• Published in: Stem cell research Vol. 28; pp. 29 - 32
• Main Authors: Tu, Jian, Huo, Zijun, Liu, Mo, Wang, Donghui, Xu, An, Zhou, Ruoji, Zhu, Dandan, Gingold, Julian, Shen, Jingnan, Zhao, Ruiying, Lee, Dung-Fang
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01.04.2018; Elsevier
• Subjects: Base Sequence; Cell Culture Techniques - methods; Cell Differentiation; Cell Line; CRISPR-Associated Protein 9 - metabolism; CRISPR-Cas Systems - genetics; Deoxyribonuclease I - metabolism; Gene Deletion; Heterozygote; Human Embryonic Stem Cells - cytology; Humans; Male; Retinoblastoma Protein - genetics
• ISSN: 1873-5061; 1876-7753; 1876-7753
• DOI: 10.1016/j.scr.2018.01.021

The Retinoblastoma 1 (RB1) tumor suppressor, a member of the Retinoblastoma gene family, functions as a pocket protein for the functional binding of E2F transcription factors. About 1/3 of retinoblastoma patients harbor a germline RB1 mutation or deletion, leading to the development of retinoblastoma. Here, we demonstrate generation of a heterozygous deletion of the RB1 gene in the H1 human embryonic stem cell line using CRISPR/Cas9 nickase genome editing. The RB1 heterozygous knockout H1 cell line shows a normal karyotype, maintains a pluripotent state, and is capable of differentiation to the three germline layers.