An Ultrasensitive Calcium Reporter System via CRISPR-Cas9-Mediated Genome Editing in Human Pluripotent Stem Cells

Yuqian Jiang; Yuxiao Zhou; Xiaoping Bao; Chuanxin Chen; Lauren N. Randolph; Jing Du; Xiaojun Lance Lian

iScience (Nov 2018)

An Ultrasensitive Calcium Reporter System via CRISPR-Cas9-Mediated Genome Editing in Human Pluripotent Stem Cells

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

Affiliations

Yuqian Jiang: Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA 16802, USA; The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA
Yuxiao Zhou: Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, University Park, PA 16802, USA
Xiaoping Bao: Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720, USA
Chuanxin Chen: Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA 16802, USA; College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China
Lauren N. Randolph: Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA 16802, USA; The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA
Jing Du: Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, University Park, PA 16802, USA
Xiaojun Lance Lian: Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA 16802, USA; The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA; Department of Biology, The Pennsylvania State University, University Park, PA 16802, USA; Corresponding author

Journal volume & issue: Vol. 9
pp. 27 – 35

Abstract

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Summary: 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. : Cell Engineering; Optical Imaging; Specialized Functions of Cells Subject Areas: Cell Engineering, Optical Imaging, Specialized Functions of Cells

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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