Generation of αMHC-EGFP knock-in in human pluripotent stem cell line, SNUe003-A-3 using CRISPR/Cas9-based gene targeting

Ha Myoung Lee; A. Hyeon Kim; Sunsook Hwang; Juwon Jung; Hyewon Seol; Jin Jea Sung; Seung Min Jeong; Young Min Choi; Jong Kwan Jun; Han Soo Kim; Jiho Jang

doi:10.1016/j.scr.2022.102779

Generation of αMHC-EGFP knock-in in human pluripotent stem cell line, SNUe003-A-3 using CRISPR/Cas9-based gene targeting

Ha Myoung Lee, A. Hyeon Kim, Sunsook Hwang, Juwon Jung, Hyewon Seol, Jin Jea Sung, Seung Min Jeong, Young Min Choi, Jong Kwan Jun, Han Soo Kim, Jiho Jang

Department of Biochemistry

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

Abstract

The cardiac muscle-specific protein, α-myosin heavy chain (αMHC), is a major component of cardiac muscle filaments involved in cardiac muscle contraction. Here, we established an αMHC-enhanced fluorescent protein (EGFP) knock-in human pluripotent stem cell (hPSC) line by linking the EGFP gene to the C-terminal region of αMHC via a 2A non-joining peptide using CRISPR/Cas9 nuclease. The EGFP reporter precisely reflected the endogenous level of αMHC upon the induction of cardiac differentiation. This reporter cell line will be a valuable platform for cardiotoxicity tests, drug screening, and investigating the pathological mechanisms of cardiomyocytes.

Original language	English
Article number	102779
Journal	Stem Cell Research
Volume	61
DOIs	https://doi.org/10.1016/j.scr.2022.102779
State	Published - May 2022

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title = "Generation of αMHC-EGFP knock-in in human pluripotent stem cell line, SNUe003-A-3 using CRISPR/Cas9-based gene targeting",

abstract = "The cardiac muscle-specific protein, α-myosin heavy chain (αMHC), is a major component of cardiac muscle filaments involved in cardiac muscle contraction. Here, we established an αMHC-enhanced fluorescent protein (EGFP) knock-in human pluripotent stem cell (hPSC) line by linking the EGFP gene to the C-terminal region of αMHC via a 2A non-joining peptide using CRISPR/Cas9 nuclease. The EGFP reporter precisely reflected the endogenous level of αMHC upon the induction of cardiac differentiation. This reporter cell line will be a valuable platform for cardiotoxicity tests, drug screening, and investigating the pathological mechanisms of cardiomyocytes.",

author = "Lee, {Ha Myoung} and Kim, {A. Hyeon} and Sunsook Hwang and Juwon Jung and Hyewon Seol and Sung, {Jin Jea} and Jeong, {Seung Min} and Choi, {Young Min} and Jun, {Jong Kwan} and Kim, {Han Soo} and Jiho Jang",

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year = "2022",

month = may,

doi = "10.1016/j.scr.2022.102779",

language = "English",

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journal = "Stem Cell Research",

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AU - Lee, Ha Myoung

AU - Kim, A. Hyeon

AU - Hwang, Sunsook

AU - Jung, Juwon

AU - Seol, Hyewon

AU - Sung, Jin Jea

AU - Jeong, Seung Min

AU - Choi, Young Min

AU - Jun, Jong Kwan

AU - Kim, Han Soo

AU - Jang, Jiho

PY - 2022/5

Y1 - 2022/5

N2 - The cardiac muscle-specific protein, α-myosin heavy chain (αMHC), is a major component of cardiac muscle filaments involved in cardiac muscle contraction. Here, we established an αMHC-enhanced fluorescent protein (EGFP) knock-in human pluripotent stem cell (hPSC) line by linking the EGFP gene to the C-terminal region of αMHC via a 2A non-joining peptide using CRISPR/Cas9 nuclease. The EGFP reporter precisely reflected the endogenous level of αMHC upon the induction of cardiac differentiation. This reporter cell line will be a valuable platform for cardiotoxicity tests, drug screening, and investigating the pathological mechanisms of cardiomyocytes.

AB - The cardiac muscle-specific protein, α-myosin heavy chain (αMHC), is a major component of cardiac muscle filaments involved in cardiac muscle contraction. Here, we established an αMHC-enhanced fluorescent protein (EGFP) knock-in human pluripotent stem cell (hPSC) line by linking the EGFP gene to the C-terminal region of αMHC via a 2A non-joining peptide using CRISPR/Cas9 nuclease. The EGFP reporter precisely reflected the endogenous level of αMHC upon the induction of cardiac differentiation. This reporter cell line will be a valuable platform for cardiotoxicity tests, drug screening, and investigating the pathological mechanisms of cardiomyocytes.

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ER -

Generation of αMHC-EGFP knock-in in human pluripotent stem cell line, SNUe003-A-3 using CRISPR/Cas9-based gene targeting

Abstract

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