An Ultrasensitive Calcium Reporter System via CRISPR-Cas9-Mediated Genome Editing in Human Pluripotent Stem Cells. Academic Article uri icon


  • Genetically encoded calcium indicator (GCaMP) proteins have been reported for imaging cardiac cell activity based on intracellular calcium transients. To bring human pluripotent stem cell (hPSC)-derived cardiomyocytes (CMs) to the clinic, it is critical to evaluate the functionality of CMs. Here, we show that GCaMP6s-expressing hPSCs can be generated and used for CM characterization. By leveraging CRISPR-Cas9 genome editing tools, we generated a knockin cell line that constitutively expresses GCaMP6s, an ultrasensitive calcium sensor protein. We further showed that this clone maintained pluripotency and cardiac differentiation potential. These knockin hPSC-derived CMs exhibited sensitive fluorescence fluctuation with spontaneous contraction. We then compared the fluorescence signal with mechanical contraction signal. The knockin hPSC-derived CMs also showed sensitive response to isoprenaline treatment in a concentration-dependent manner. Therefore, the GCaMP6s knockin hPSC line provides a non-invasive, sensitive, and economic approach to characterize the functionality of hPSC-derived CMs.

published proceedings

  • iScience

altmetric score

  • 41.5

author list (cited authors)

  • Jiang, Y., Zhou, Y., Bao, X., Chen, C., Randolph, L. N., Du, J., & Lian, X. L.

citation count

  • 16

complete list of authors

  • Jiang, Yuqian||Zhou, Yuxiao||Bao, Xiaoping||Chen, Chuanxin||Randolph, Lauren N||Du, Jing||Lian, Xiaojun Lance

publication date

  • November 2018