Department of Oncology, Hematology, Immunology, and Rheumatology, University Hospital Tuebingen, Tuebingen, Germany.
Methods Mol Biol. 2020;2115:471-483. doi: 10.1007/978-1-0716-0290-4_27.
Research on patient-derived induced pluripotent stem cells (iPSCs) could immensely benefit from the implementation of CRISPR/Cas9 genome editing of iPSCs, creating unique opportunities such as the establishment of isogenic iPSC lines for disease modeling or personalized patient-specific drug screenings. Here we describe a stepwise protocol of safe, efficient, and selection-free CRISPR/Cas9-mediated gene correction or knockout in human iPSCs followed by 3D spin-embryoid body (EB)-based hematopoietic/neutrophilic iPSC-differentiation.
利用 CRISPR/Cas9 基因组编辑技术对患者来源的诱导多能干细胞（iPSC）进行研究，可以极大地受益，为疾病建模或个性化患者特异性药物筛选等提供建立同基因 iPSC 系的独特机会。在此，我们描述了一种安全、高效且无需选择的 CRISPR/Cas9 介导的基因校正或敲除的分步方案，随后进行基于 3D 旋转胚胎体（EB）的造血/中性粒细胞 iPSC 分化。
