Establishment of TUBB3-mCherry knock-in human pluripotent stem cell line using CRISPR/Cas9 (SNUe003-A-4)

A-Hyeon Kim; Ha Myoung Lee; Hong-Sik Kim; Juwon Jung; Hyewon Seol; Eunmi Choi; Seongeun Lee; Young Min Choi; Jong Kwan Jun; Han-Soo Kim; Jiho Jang

Stem Cell Research (Jun 2023)

Establishment of TUBB3-mCherry knock-in human pluripotent stem cell line using CRISPR/Cas9 (SNUe003-A-4)

A-Hyeon Kim,
Ha Myoung Lee,
Hong-Sik Kim,
Juwon Jung,
Hyewon Seol,
Eunmi Choi,
Seongeun Lee,
Young Min Choi,
Jong Kwan Jun,
Han-Soo Kim,
Jiho Jang

Affiliations

A-Hyeon Kim: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea
Ha Myoung Lee: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea
Hong-Sik Kim: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea
Juwon Jung: Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
Hyewon Seol: Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
Eunmi Choi: R&D Center, RADIANT Inc., 31, Geodudanji 1 gil, Dongnae-myeon, Chuncheon, Gangwon 24398, Republic of Korea
Seongeun Lee: R&D Center, RADIANT Inc., 31, Geodudanji 1 gil, Dongnae-myeon, Chuncheon, Gangwon 24398, Republic of Korea
Young Min Choi: Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
Jong Kwan Jun: Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
Han-Soo Kim: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea; Corresponding authors.
Jiho Jang: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea; Corresponding authors.

Journal volume & issue: Vol. 69
p. 103064

Abstract

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TUBB3 is a structural neuronal protein important for multiple neuronal functions including axonal guidance and maturation. This study aimed to generate a human pluripotent stem cell (hPSC) line with a TUBB3-mCherry reporter using CRISPR/SpCas9 nuclease. The stop codon in the last exon of TUBB3 was replaced with a T2A-mCherry cassette using CRISPR/SpCas9-mediated homologous recombination. The established TUBB3-mCherry knock-in cell line exhibited typical pluripotent characteristics. The mCherry reporter faithfully replicated the endogenous level of TUBB3 upon induction of neuronal differentiation. The reporter cell line could contribute to the investigation of neuronal differentiation, neuronal toxicity, and neuronal tracing.

Published in Stem Cell Research

ISSN: 1873-5061 (Print); 1876-7753 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Science: Biology (General)
Website: https://www.journals.elsevier.com/stem-cell-research

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