RRC ID 81693
Author Takagi D, Tsukamoto S, Nakade K, Shimizu T, Arai Y, Matsuo-Takasaki M, Noguchi M, Nakamura Y, Yumoto N, Kawada J, Hayata T, Hayashi Y.
Title Generation of MBP-tdTomato reporter human induced pluripotent stem cell line for live myelin visualization.
Journal Stem Cell Res
Abstract Myelin basic protein (MBP) is a major component of the myelin sheaths of oligodendrocytes in the central nervous system and Schwann cells of the peripheral nervous system. Here we generated heterozygous fluorescent reporter of MBP gene in human induced pluripotent stem cells (hiPSCs). CRISPR/Cas9 genome editing technology was employed to knock in fused tdTomato fluorescent protein and EF1 alpha promoter-driven Bleomycin (Zeocin) resistance gene to the translational MBP C-terminal region. The resulting line, MBP-TEZ, showed tdTomato fluorescence upon oligodendrocyte differentiation. This reporter hiPSC line provides a precedential opportunity for monitoring human myelin formation and degeneration and purifying MBP-expressing cell lineages.
Volume 79
Pages 103493
Published 2024-9-1
DOI 10.1016/j.scr.2024.103493
PII S1873-5061(24)00191-0
PMID 39032428
MeSH CRISPR-Cas Systems Cell Differentiation Cell Line Gene Editing Genes, Reporter Humans Induced Pluripotent Stem Cells* / cytology Induced Pluripotent Stem Cells* / metabolism Luminescent Proteins / genetics Luminescent Proteins / metabolism Myelin Basic Protein / genetics Myelin Basic Protein / metabolism Myelin Sheath / metabolism Red Fluorescent Protein
IF 4.495
Resource
DNA material pUCtk2x2 (RDB19569) pX330-MBP (RDB20337) pUCtk2X2NN_MBP-2xLG3-TEZ (RDB20338)
Human and Animal Cells 1383D6(HPS1006)