Base Editing Mediated Generation of Point Mutations Into Human Pluripotent Stem Cells for Modeling Disease

• Published in: Frontiers in cell and developmental biology 2020-09, Vol.8, p.590581-590581
• Main Authors: Qi, Tao, Wu, Fujian, Xie, Yuquan, Gao, Siqi, Li, Miaomiao, Pu, Jun, Li, Dali, Lan, Feng, Wang, Yongming
• Format: Article
• Language: English
• Subjects: base editing; Cell and Developmental Biology; disease modeling; episomal vector; human pluripotent stem cell; IPS; long QT syndrome
• ISSN: 2296-634X; 2296-634X
• DOI: 10.3389/fcell.2020.590581

Human pluripotent stem cells (hPSCs) are a powerful platform for disease modeling and drug discovery. However, the introduction of known pathogenic mutations into hPSCs is a time-consuming and labor-intensive process. Base editing is a newly developed technology that enables facile introduction of point mutations into specific loci within the genome of living cells. Here, we design an all-in-one episomal vector that expresses a single guide RNA (sgRNA) with an adenine base editor (ABE) or a cytosine base editor (CBE). Both ABE and CBE can efficiently introduce mutations into cells, A-to-G and C-to-T, respectively. We introduce disease-specific mutations of long QT syndrome into hPSCs to model LQT1, LQT2, and LQT3. Electrophysiological analysis of hPSC-derived cardiomyocytes (hPSC-CMs) using multi-electrode arrays (MEAs) reveals that edited hPSC-CMs display significant increases in duration of the action potential. Finally, we introduce the novel Brugada syndrome-associated mutation into hPSCs, demonstrating that this mutation can cause abnormal electrophysiology. Our study demonstrates that episomal encoded base editors (epi-BEs) can efficiently generate mutation-specific disease hPSC models.