Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1

Yi-Hung Chen; Pei-San Huang; Meng-Hsuan Wen; Manhua Pan; Dung-Fang Lee; Tai-Yen Chen

doi:10.1016/j.scr.2022.102845

Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1

Stem Cell Res. 2022 Aug:63:102845. doi: 10.1016/j.scr.2022.102845. Epub 2022 Jun 14.

Authors

Yi-Hung Chen¹, Pei-San Huang², Meng-Hsuan Wen², Manhua Pan², Dung-Fang Lee³, Tai-Yen Chen⁴

Affiliations

¹ Department of Chemistry, University of Houston, Houston, TX 77204, USA; Development, Disease Models and Therapeutics Graduate Program, Baylor College of Medicine, Houston, TX 77030, USA.
² Department of Chemistry, University of Houston, Houston, TX 77204, USA.
³ Department of Integrative Biology and Pharmacology, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, TX 77030, USA; Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Center for Precision Health, School of Biomedical Informatics, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA.
⁴ Department of Chemistry, University of Houston, Houston, TX 77204, USA. Electronic address: tchen37@central.uh.edu.

Abstract

Copper transporter 1 (CTR1) is the major membrane protein responsible for cellular copper (Cu) uptake and mediates cellular copper homeostasis. To elucidate CTR1's behavior using imaging approaches, we generated a homozygous knock-in human embryonic stem cell (hESC) clone expressing photoconvertible fluorescence protein mEos4b-tagged endogenous CTR1 using CRISPR-Cas9 mediated homologous recombination. The engineered cells express functional CTR1-mEos4b fusion and have normal stem cell morphology. They remain pluripotent and can be differentiated into all three germ layers in vitro. This resource allows the study of CTR1 at an endogenous level in different cellular contexts using microscopy.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Cation Transport Proteins* / genetics
Cation Transport Proteins* / metabolism
Cell Line
Copper / metabolism
Copper Transporter 1
Homozygote
Human Embryonic Stem Cells* / metabolism
Humans

Substances

Cation Transport Proteins
Copper Transporter 1
Copper

Grants and funding

R35 GM133505/GM/NIGMS NIH HHS/United States