Generation of heterozygous (MRli003-A-5) and homozygous (MRli003-A-6) voltage-sensing knock-in human iPSC lines by CRISPR/Cas9 editing of the AAVS1 locus

Assessment of the electrophysiological properties of cardiomyocytes is necessary for phenotyping cardiac disorders and for drug screening. Optical action potential imaging using a genetically encoded voltage-sensing fluorescent protein (VSFP) allows for high-throughput functional characterization of cardiomyocytes, which offers an advantage over the traditional patch-clamp technique. Here, we knocked VSFP into the AAVS1 safe harbor locus of human iPSCs, generating two stable voltage indicator lines -one heterozygous (MRIi003-A-5) and the other homozygous (MRI003-A-6). Both lines can be used for optical membrane potential recordings and provide a powerful platform for a wide range of applications in cardiovascular biomedicine.

Automated summary

Assessment of cardiomyocyte electrophysiological properties is crucial for understanding cardiac disorders and drug screening. Optical action potential imaging using a genetically encoded voltage-sensing fluorescent protein (VSFP) provides a high-throughput method for functional characterization of cardiomyocytes. This study successfully integrated VSFP into human induced pluripotent stem cells, creating two stable voltage indicator lines, which can be used for optical membrane potential recordings and have wide applications in cardiovascular biomedicine.