iPS Cell Advanced Characterization and Development Team, BioResource Research Center, RIKEN, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan; Department of Molecular Pharmacology, Graduate School of Pharmaceutical Sciences and Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
iPS Cell Advanced Characterization and Development Team, BioResource Research Center, RIKEN, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.
Stem Cell Res. 2024 Sep;79:103493. doi: 10.1016/j.scr.2024.103493. Epub 2024 Jul 11.
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.
髓鞘碱性蛋白（MBP）是中枢神经系统少突胶质细胞和周围神经系统雪旺细胞髓鞘的主要成分。在这里，我们在人诱导多能干细胞（hiPSC）中生成了 MBP 基因的杂合荧光报告基因。CRISPR/Cas9 基因组编辑技术被用来将融合的 tdTomato 荧光蛋白和 EF1α启动子驱动的博来霉素（zeocin）抗性基因敲入到 MBP C 端区域的翻译区。所得的 MBP-TEZ 系在少突胶质细胞分化时显示 tdTomato 荧光。该报告基因 hiPSC 系为监测人髓鞘形成和退化以及纯化 MBP 表达细胞谱系提供了一个先例机会。
