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

• Main Authors: Tao Qi, Fujian Wu, Yuquan Xie, Siqi Gao, Miaomiao Li, Jun Pu, Dali Li, Feng Lan, Yongming Wang
• Format: Article
• Language: English
• Published: Frontiers Media S.A. 2020-09-01
• Series: Frontiers in Cell and Developmental Biology
• Subjects: human pluripotent stem cell; base editing; episomal vector; disease modeling; IPS; long QT syndrome
• Online Access: https://www.frontiersin.org/article/10.3389/fcell.2020.590581/full

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.