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Establishment of TUBB3-mCherry knock-in human pluripotent stem cell line using CRISPR/Cas9 (SNUe003-A-4).
Kim, A-Hyeon; Lee, Ha Myoung; Kim, Hong-Sik; Jung, Juwon; Seol, Hyewon; Choi, Eunmi; Lee, Seongeun; Min Choi, Young; Jun, Jong Kwan; Kim, Han-Soo; Jang, Jiho.
Affiliation
  • Kim AH; Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea.
  • Lee HM; Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea.
  • Kim HS; Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea.
  • Jung J; Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
  • Seol H; Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
  • Choi E; R&D Center, RADIANT Inc., 31, Geodudanji 1 gil, Dongnae-myeon, Chuncheon, Gangwon 24398, Republic of Korea.
  • Lee S; R&D Center, RADIANT Inc., 31, Geodudanji 1 gil, Dongnae-myeon, Chuncheon, Gangwon 24398, Republic of Korea.
  • Min Choi Y; Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
  • Jun JK; Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
  • Kim HS; Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea. Electronic address: hansk@cku.ac.kr.
  • Jang J; Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea. Electronic address: jhjang@cku.ac.kr.
Stem Cell Res ; 69: 103064, 2023 06.
Article in En | MEDLINE | ID: mdl-36913849
ABSTRACT
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.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Pluripotent Stem Cells / CRISPR-Cas Systems Limits: Humans Language: En Journal: Stem Cell Res Year: 2023 Document type: Article
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Pluripotent Stem Cells / CRISPR-Cas Systems Limits: Humans Language: En Journal: Stem Cell Res Year: 2023 Document type: Article