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

Fangfang Zhang; Anna B. Meier; Daniel Sinnecker; Stefan Engelhardt; Peter Lipp; Karl Ludwig Laugwitz; Tatjana Dorn; Alessandra Moretti

doi:10.1016/j.scr.2022.102785

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

Fangfang Zhang
, Anna B. Meier
, Daniel Sinnecker
, Stefan Engelhardt
, Peter Lipp
, Karl Ludwig Laugwitz
, Tatjana Dorn
, Alessandra Moretti

Technical University of Munich
Partner Site Munich Heart Alliance
Saarland University Medical Center

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

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.

Original language	English
Article number	102785
Journal	Stem Cell Research
Volume	61
DOIs	https://doi.org/10.1016/j.scr.2022.102785
State	Published - May 2022

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title = "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",

abstract = "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.",

author = "Fangfang Zhang and Meier, \{Anna B.\} and Daniel Sinnecker and Stefan Engelhardt and Peter Lipp and Laugwitz, \{Karl Ludwig\} and Tatjana Dorn and Alessandra Moretti",

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year = "2022",

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doi = "10.1016/j.scr.2022.102785",

language = "English",

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AU - Zhang, Fangfang

AU - Meier, Anna B.

AU - Sinnecker, Daniel

AU - Engelhardt, Stefan

AU - Lipp, Peter

AU - Laugwitz, Karl Ludwig

AU - Dorn, Tatjana

AU - Moretti, Alessandra

PY - 2022/5

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AB - 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.

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AN - SCOPUS:85127860042

SN - 1873-5061

VL - 61

JO - Stem Cell Research

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ER -

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

Abstract

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