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

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...

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Autores principales: Chen, Yi-Hung, Huang, Pei-San, Wen, Meng-Hsuan, Pan, Manhua, Lee, Dung-Fang, Chen, Tai-Yen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2022
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9360979/
https://www.ncbi.nlm.nih.gov/pubmed/35728441
http://dx.doi.org/10.1016/j.scr.2022.102845
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author Chen, Yi-Hung
Huang, Pei-San
Wen, Meng-Hsuan
Pan, Manhua
Lee, Dung-Fang
Chen, Tai-Yen
author_facet Chen, Yi-Hung
Huang, Pei-San
Wen, Meng-Hsuan
Pan, Manhua
Lee, Dung-Fang
Chen, Tai-Yen
author_sort Chen, Yi-Hung
collection PubMed
description 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.
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spelling pubmed-93609792022-08-09 Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1 Chen, Yi-Hung Huang, Pei-San Wen, Meng-Hsuan Pan, Manhua Lee, Dung-Fang Chen, Tai-Yen Stem Cell Res Article 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. 2022-08 2022-06-14 /pmc/articles/PMC9360979/ /pubmed/35728441 http://dx.doi.org/10.1016/j.scr.2022.102845 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) ).
spellingShingle Article
Chen, Yi-Hung
Huang, Pei-San
Wen, Meng-Hsuan
Pan, Manhua
Lee, Dung-Fang
Chen, Tai-Yen
Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1
title Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1
title_full Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1
title_fullStr Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1
title_full_unstemmed Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1
title_short Generation of a homozygous knock-in human embryonic stem cell line expressing mEos4b-tagged CTR1
title_sort generation of a homozygous knock-in human embryonic stem cell line expressing meos4b-tagged ctr1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9360979/
https://www.ncbi.nlm.nih.gov/pubmed/35728441
http://dx.doi.org/10.1016/j.scr.2022.102845
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