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Generation of MBP-tdTomato reporter human induced pluripotent stem cell line for live myelin visualization
oleh: Daigo Takagi, Satomi Tsukamoto, Koji Nakade, Tomoya Shimizu, Yutaka Arai, Mami Matsuo-Takasaki, Michiya Noguchi, Yukio Nakamura, Norihiro Yumoto, Jiro Kawada, Tadayoshi Hayata, Yohei Hayashi
Format: | Article |
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Diterbitkan: | Elsevier 2024-09-01 |
Deskripsi
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.