The establishment of a homozygous SNTA1 knockout human embryonic stem cell line (WAe009-A-50) using the CRISPR/Cas9 system

Dong, Tao; Zhang, Siyao; Chang, Yun; Bai, Rui; Jiang, Yuliang; Ma, Shuhong; Li, Yanan; Jiang, Hong; Lü, Wenjing

doi:10.1016/j.scr.2021.102196

Cited by 5 publications

(5 citation statements)

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“…Furthermore, western blotting confirmed that H9SN-TA1KO is SNTA1 deficient (Additional file 2: Fig. 1C) [27].…”

Section: Establishment Of Homozygous Snta1-deficient Hescs (H9snta1ko)mentioning

confidence: 73%

“…Further study on patients' iPSCs was limited by different genetic backgrounds. With the widespread applications of gene-editing technologies in eukaryotic cells [23][24][25], the formation of genetically edited human embryonic stem cells can help overcome the barrier of genetic diversity [26,27].Using the CRISPR-Cas9 system we established the H9SN-TA1KO from the H9 embryonic stem cells, and then SNTA1-deficient cardiomyocytes were inducted from the H9SNTA1KO. The phenotype of SNTA1-deficient cardiomyocytes was investigated.…”

Section: Introductionmentioning

confidence: 99%

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SNTA1-deficient human cardiomyocytes demonstrate hypertrophic phenotype and calcium handling disorder

et al. 2022

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Background α-1-syntrophin (SNTA1), a protein encoded by SNTA1, is highly expressed in human cardiomyocytes. Mutations in SNTA1 are associated with arrhythmia and cardiomyopathy. Previous research on SNTA1 has been based on non-human cardiomyocytes. This study was designed to identify the phenotype of SNTA1-deficiency using human cardiomyocytes. Methods SNTA1 was knocked out in the H9 embryonic stem cell line using the CRISPR-Cas9 system. H9SNTA1KO cells were then induced to differentiate into cardiomyocytes using small molecule inhibitors. The phenotypic discrepancies associated with SNTA1-deficient cardiomyocytes were investigated. Results SNTA1 was truncated at the 149th amino acid position of PH1 domain by a stop codon (TGA) using the CRISPR-Cas9 system. SNTA1-deficiency did not affect the pluripotency of H9SNTA1KO, and they retain their in vitro ability to differentiate into cardiomyocytes. However, H9SNTA1KO derived cardiomyocytes exhibited hypertrophic phenotype, lower cardiac contractility, weak calcium transient intensity, and lower level of calcium in the sarcoplasmic reticulum. Early treatment of SNTA1-deficient cardiomyocytes with ranolazine improved the calcium transient intensity and cardiac contractility. Conclusion SNTA1-deficient cardiomyocytes can be used to research the etiology, pathogenesis, and potential therapies for myocardial diseases. The SNTA1-deficient cardiomyocyte model suggests that the maintenance of cardiac calcium homeostasis is a key target in the treatment of myocardial-related diseases.

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“…Furthermore, western blotting confirmed that H9SN-TA1KO is SNTA1 deficient (Additional file 2: Fig. 1C) [27].…”

Section: Establishment Of Homozygous Snta1-deficient Hescs (H9snta1ko)mentioning

confidence: 73%

Section: Introductionmentioning

confidence: 99%

SNTA1-deficient human cardiomyocytes demonstrate hypertrophic phenotype and calcium handling disorder

et al. 2022

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“…It has been confirmed that the vertebrae of sheep and humans are most similar in the thoracic and lumbar regions, although they show substantial differences in some dimensions ( 33 ). SNTA1 encodes the α1-synthetic protein, a scaffold protein that is a component of the anti-muscular dystrophin-associated protein complex ( 34 ). SNTA1 is the link between the extracellular matrix, the intracellular signaling apparatus, and the actin cytoskeleton.…”

Section: Discussionmentioning

confidence: 99%

Effects of nonsynonymous single nucleotide polymorphisms of the KIAA1217, SNTA1 and LTBP1 genes on the growth traits of Ujumqin sheep

Liu,

Qin,

Zhang

et al. 2024

Front. Vet. Sci.

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Sheep body size can directly reflect the growth rates and fattening rates of sheep and is also an important index for measuring the growth performance of meat sheep. In this study, high-resolution resequencing data from four sheep breeds (Dorper sheep, Suffolk sheep, Ouessant sheep, and Shetland sheep) were analyzed. The nonsynonymous single nucleotide polymorphisms of three candidate genes (KIAA1217, SNTA1, and LTBP1) were also genotyped in 642 healthy Ujumqin sheep using MALDI-TOFMS and the genotyping results were associated with growth traits. The results showed that different genotypes of the KIAA1217 g.24429511T>C locus had significant effects on the chest circumferences of Ujumqin sheep. The SNTA1 g.62222626C>A locus had different effects on the chest depths, shoulder widths and rump widths of Ujumqin sheep. This study showed that these two sites can be used for marker-assisted selection, which will be beneficial for future precision molecular breeding.

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“…There are over 15 genes involved in causing LQTS, which usually are either encoding ion channels or associated proteins ( Ueda et al, 2008 ), and among them we can find SNTA1 ( Mizusawa et al, 2014 ). SNTA1 encodes α1-syntrophin, a scaffold constituent protein of the dystrophin-associated protein complex ( Dong et al, 2021 ). Therefore, cardiac models generated from human iPSCs can help to understand better the functional consequences of the different mutations implicated in LQTS patients ( Wu et al, 2019 ).…”

Section: Resource Detailsmentioning

confidence: 99%

Generation of two iPSC lines from long QT syndrome patients carrying SNTA1 variants

et al. 2023

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The establishment of a homozygous SNTA1 knockout human embryonic stem cell line (WAe009-A-50) using the CRISPR/Cas9 system

Cited by 5 publications

References 1 publication

SNTA1-deficient human cardiomyocytes demonstrate hypertrophic phenotype and calcium handling disorder

SNTA1-deficient human cardiomyocytes demonstrate hypertrophic phenotype and calcium handling disorder

Effects of nonsynonymous single nucleotide polymorphisms of the KIAA1217, SNTA1 and LTBP1 genes on the growth traits of Ujumqin sheep

Generation of two iPSC lines from long QT syndrome patients carrying SNTA1 variants

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