Establishment of two iPSC lines from healthy donor with heterozygous mutation in the SLC26A4 gene

Li, Sijun; He, Chufeng; Qi, Feng; Bai, Yijiang; Liu, Xianlin; Ling, Jie; Mei, Lingyun; Wu, Xuewen; Song, Jian

doi:10.1016/j.scr.2022.102916

Cited by 3 publications

(4 citation statements)

References 2 publications

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“…The human peripheral blood mononuclear cells (PBMCs) -derived iPSCs used in this study were generated from two patients with clinical phenotypes con rmed OI. The establishment of OI -iPSCs from OI-PBMCs was performed according to a method previously reported under feeder-and serum-free conditions 16,17 . Culturing PBMCs were transfected with episomal vectors 18 on Nucleofector™ 2b device (Lonza) using Kit VCA-1003 (Lonza) 17 .…”

Section: Establishment and Characterization Of Oi-pbmc-derived Oi-ipscmentioning

confidence: 99%

“…The establishment of OI -iPSCs from OI-PBMCs was performed according to a method previously reported under feeder-and serum-free conditions 16,17 . Culturing PBMCs were transfected with episomal vectors 18 on Nucleofector™ 2b device (Lonza) using Kit VCA-1003 (Lonza) 17 . After electroporation, the cells were seeded in a 12-well plate and cultured in WiCell + VitC medium.…”

Section: Establishment and Characterization Of Oi-pbmc-derived Oi-ipscmentioning

confidence: 99%

“…The CRISPR-Cas9 machinery was delivered to iPSCs and analyzed according to previously published methods 17 . Brie y, sg4-spCas9 and HDR plasmids were transfected at a 1:2 molar ratio into patient iPSCs using the Human Stem Cell Nucleofector® Kit 1 (Lonza, No.…”

Section: Cloning Of Crispr-cas9 and Hdr Donor Constructmentioning

confidence: 99%

“…Reprogramming of induced pluripotent stem cells (iPSCs) was performed following a previously published protocol 16,17 (Fig. 3A).…”

Section: Generation Of Induced Pluripotent Stem Cells (Ipscs) From Oi...mentioning

confidence: 99%

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Identification of a family with van der Hoeve’s syndrome harboring a novel COL1A1 mutation and generation of patient-derived iPSC lines and CRISPR/Cas9-corrected isogenic iPSCs

Li,

Mei,

et al. 2024

Human Cell

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Van der Hoeve's syndrome, also known as osteogenesis imperfecta (OI), is a genetic connective tissue disorder characterized by fragile, fracture-prone bone and hearing loss. The disease is caused by a gene mutation in one of the two type I collagen genes COL1A1 or COL1A2. In this study, we identi ed a novel frameshift mutation of the COL1A1 gene (c.1607delG) in a family with OI using whole-exome sequencing, bioinformatics analysis and Sanger sequencing. This mutation may lead to the deletion of a portion of exon 23 and the generation of a premature stop codon in the COL1A1 gene. To further investigate the impact of this mutation, we established two induced pluripotent stem cell (iPSC) lines from peripheral blood mononuclear cells (PBMC) of OI patients carrying a novel mutation in the COL1A1 gene. Using a CRISPR-based homology-directed repair strategy, we corrected the OI disease-causing COL1A1 novel mutations in iPSCs generated from an affected individual. Two OI patients' iPSCs and corrected OI-iPSC lines displayed a normal karyotype, morphology, pluripotency, and potential to differentiate into three germ layers. These ndings demonstrate the potential of iPSCs derived from affected OI patients and corrected OI-iPSCs as a tool to study the pathophysiologic mechanisms of speci c mutations. Overall, our results provide new insights into the genetic basis of Van der Hoeve's syndrome and highlight the potential of iPSC technology for disease modeling and therapeutic development.

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Section: Establishment and Characterization Of Oi-pbmc-derived Oi-ipscmentioning

confidence: 99%

Section: Establishment and Characterization Of Oi-pbmc-derived Oi-ipscmentioning

confidence: 99%

Section: Cloning Of Crispr-cas9 and Hdr Donor Constructmentioning

confidence: 99%

“…Reprogramming of induced pluripotent stem cells (iPSCs) was performed following a previously published protocol 16,17 (Fig. 3A).…”

Section: Generation Of Induced Pluripotent Stem Cells (Ipscs) From Oi...mentioning

confidence: 99%

See 2 more Smart Citations

Identification of a family with van der Hoeve’s syndrome harboring a novel COL1A1 mutation and generation of patient-derived iPSC lines and CRISPR/Cas9-corrected isogenic iPSCs

Li,

Mei,

et al. 2024

Human Cell

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PAX3 mutation suppress otic progenitors proliferation and induce apoptosis by inhibiting WNT1/β-catenin signaling pathway in WS1 patient iPSC-derived inner ear organoids

Li,

He,

Mei

et al. 2024

Biochemical and Biophysical Research Communications

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Identification of a Family with van der Hoeve's syndrome Harboring a Novel COL1A1 Mutation and Generation of Patient-Derived iPSC Lines and CRISPR/Cas9-Corrected Isogenic iPSCs

Li,

Mei,

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Van der Hoeve's syndrome, also known as osteogenesis imperfecta (OI), is a genetic connective tissue disorder characterized by fragile, fracture-prone bone and hearing loss. The disease is caused by a gene mutation in one of the two type I collagen genes COL1A1 or COL1A2. In this study, we identified a novel frameshift mutation of the COL1A1 gene (c.1607delG) in a family with OI using whole-exome sequencing, bioinformatics analysis and Sanger sequencing. This mutation may lead to the deletion of a portion of exon 23 and the generation of a premature stop codon in the COL1A1 gene. To further investigate the impact of this mutation, we established two induced pluripotent stem cell (iPSC) lines from peripheral blood mononuclear cells (PBMC) of OI patients carrying a novel mutation in the COL1A1 gene. Using a CRISPR-based homology-directed repair strategy, we corrected the OI disease-causing COL1A1 novel mutations in iPSCs generated from an affected individual. Two OI patients' iPSCs and corrected OI-iPSC lines displayed a normal karyotype, morphology, pluripotency, and potential to differentiate into three germ layers. These findings demonstrate the potential of iPSCs derived from affected OI patients and corrected OI-iPSCs as a tool to study the pathophysiologic mechanisms of specific mutations. Overall, our results provide new insights into the genetic basis of Van der Hoeve's syndrome and highlight the potential of iPSC technology for disease modeling and therapeutic development.

show abstract

Establishment of two iPSC lines from healthy donor with heterozygous mutation in the SLC26A4 gene

Cited by 3 publications

References 2 publications

Identification of a family with van der Hoeve’s syndrome harboring a novel COL1A1 mutation and generation of patient-derived iPSC lines and CRISPR/Cas9-corrected isogenic iPSCs

Identification of a family with van der Hoeve’s syndrome harboring a novel COL1A1 mutation and generation of patient-derived iPSC lines and CRISPR/Cas9-corrected isogenic iPSCs

PAX3 mutation suppress otic progenitors proliferation and induce apoptosis by inhibiting WNT1/β-catenin signaling pathway in WS1 patient iPSC-derived inner ear organoids

Identification of a Family with van der Hoeve's syndrome Harboring a Novel COL1A1 Mutation and Generation of Patient-Derived iPSC Lines and CRISPR/Cas9-Corrected Isogenic iPSCs

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