Generation of a FTO gene knockout human embryonic stem cell line using CRISPR/Cas9 editing

Cong Wei; Qian Luo; Binsheng Wang; Yan Long; Meng Zhang; Wei Shan; Xiaohong Yu; Yulin Xu; Pengxu Qian; He Huang

doi:10.1016/j.scr.2021.102362

Generation of a FTO gene knockout human embryonic stem cell line using CRISPR/Cas9 editing

Stem Cell Res. 2021 May:53:102362. doi: 10.1016/j.scr.2021.102362. Epub 2021 Apr 22.

Authors

Cong Wei¹, Qian Luo¹, Binsheng Wang¹, Yan Long¹, Meng Zhang¹, Wei Shan¹, Xiaohong Yu¹, Yulin Xu¹, Pengxu Qian², He Huang³

Affiliations

¹ Bone Marrow Transplantation Center, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310012, Zhejiang, PR China; Institute of Hematology, Zhejiang University, Hangzhou 310012, Zhejiang, PR China.
² Bone Marrow Transplantation Center, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310012, Zhejiang, PR China; Institute of Hematology, Zhejiang University, Hangzhou 310012, Zhejiang, PR China; Zhejiang Province Engineering Laboratory for Stem Cell and Immunity Therapy, Hangzhou 310012, Zhejiang, PR China; Liangzhu Laboratory, Zhejiang University Medical Center, 1369 West Wenyi Road, Hangzhou 311121, PR China. Electronic address: axu@zju.edu.cn.
³ Bone Marrow Transplantation Center, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310012, Zhejiang, PR China; Institute of Hematology, Zhejiang University, Hangzhou 310012, Zhejiang, PR China; Zhejiang Province Engineering Laboratory for Stem Cell and Immunity Therapy, Hangzhou 310012, Zhejiang, PR China; Liangzhu Laboratory, Zhejiang University Medical Center, 1369 West Wenyi Road, Hangzhou 311121, PR China. Electronic address: huanghe@zju.edu.cn.

PMID: 34087991
DOI: 10.1016/j.scr.2021.102362

Abstract

Fat mass and obesity-associated protein (FTO) is the first protein found to have the activity of N6-methyladenosine (m6A) demethylation. It has been reported that FTO was involved in different physiological and pathological processes, including stem cell differentiation, sex determination, tumorigenesis, and progression. To further understand the exact role of FTO in these processes, we generated a FTO knockout human embryonic stem cell (hESC) line by CRISPR/Cas9 mediated gene editing method. This cell line maintained normal karyotype, pluripotency, and trilineage differentiation potential, which are considered as a model for function studies of the FTO protein in hESC self-renewal and differentiation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alpha-Ketoglutarate-Dependent Dioxygenase FTO / genetics
CRISPR-Cas Systems* / genetics
Cell Line
Gene Editing
Gene Knockout Techniques
Human Embryonic Stem Cells*
Humans

Substances

Alpha-Ketoglutarate-Dependent Dioxygenase FTO
FTO protein, human