CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L)
To produce in vitro models of human chondrodysplasias caused by dominant missense mutations in TRPV4, we used CRISPR/Cas9 gene editing to introduce two heterozygous patient mutations (p.F273L and p.P799L) into an established control human iPSC line. This control line expressed a fluorescent reporter...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-10-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506120302439 |
| _version_ | 1828465093244878848 |
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| author | Yudha Nur Patria Tayla Stenta Jinia Lilianty Lynn Rowley Edouard G. Stanley Andrew G. Elefanty John F. Bateman Shireen R. Lamandé |
| author_facet | Yudha Nur Patria Tayla Stenta Jinia Lilianty Lynn Rowley Edouard G. Stanley Andrew G. Elefanty John F. Bateman Shireen R. Lamandé |
| author_sort | Yudha Nur Patria |
| collection | DOAJ |
| description | To produce in vitro models of human chondrodysplasias caused by dominant missense mutations in TRPV4, we used CRISPR/Cas9 gene editing to introduce two heterozygous patient mutations (p.F273L and p.P799L) into an established control human iPSC line. This control line expressed a fluorescent reporter (tdTomato) at the SOX9 locus to allow real-time monitoring of cartilage differentiation by SOX9 expression. Both TRPV4 mutant iPSC lines had normal karyotypes, expressed pluripotency markers, and could differentiate into cells representative of the three embryonic germ layers. These iPSC lines, with the parental isogenic control, will be used to study TRPV4 chondrodysplasia mechanisms and explore therapeutic approaches. |
| first_indexed | 2024-12-11T03:26:17Z |
| format | Article |
| id | doaj.art-1b30b9d66478485f850dcb65bca8ce26 |
| institution | Directory Open Access Journal |
| issn | 1873-5061 |
| language | English |
| last_indexed | 2024-12-11T03:26:17Z |
| publishDate | 2020-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-1b30b9d66478485f850dcb65bca8ce262022-12-22T01:22:29ZengElsevierStem Cell Research1873-50612020-10-0148101942CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L)Yudha Nur Patria0Tayla Stenta1Jinia Lilianty2Lynn Rowley3Edouard G. Stanley4Andrew G. Elefanty5John F. Bateman6Shireen R. Lamandé7Murdoch Children’s Research Institute, Australia; Department of Paediatrics, University of Melbourne, Australia; Department of Child Health, Universitas Gadjah Mada, IndonesiaMurdoch Children’s Research Institute, AustraliaMurdoch Children’s Research Institute, Australia; Department of Paediatrics, University of Melbourne, AustraliaMurdoch Children’s Research Institute, AustraliaMurdoch Children’s Research Institute, Australia; Department of Paediatrics, University of Melbourne, Australia; Department of Anatomy and Developmental Biology, Monash University, AustraliaMurdoch Children’s Research Institute, Australia; Department of Paediatrics, University of Melbourne, Australia; Department of Anatomy and Developmental Biology, Monash University, AustraliaMurdoch Children’s Research Institute, Australia; Department of Paediatrics, University of Melbourne, Australia; Corresponding author.Murdoch Children’s Research Institute, Australia; Department of Paediatrics, University of Melbourne, AustraliaTo produce in vitro models of human chondrodysplasias caused by dominant missense mutations in TRPV4, we used CRISPR/Cas9 gene editing to introduce two heterozygous patient mutations (p.F273L and p.P799L) into an established control human iPSC line. This control line expressed a fluorescent reporter (tdTomato) at the SOX9 locus to allow real-time monitoring of cartilage differentiation by SOX9 expression. Both TRPV4 mutant iPSC lines had normal karyotypes, expressed pluripotency markers, and could differentiate into cells representative of the three embryonic germ layers. These iPSC lines, with the parental isogenic control, will be used to study TRPV4 chondrodysplasia mechanisms and explore therapeutic approaches.http://www.sciencedirect.com/science/article/pii/S1873506120302439 |
| spellingShingle | Yudha Nur Patria Tayla Stenta Jinia Lilianty Lynn Rowley Edouard G. Stanley Andrew G. Elefanty John F. Bateman Shireen R. Lamandé CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L) Stem Cell Research |
| title | CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L) |
| title_full | CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L) |
| title_fullStr | CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L) |
| title_full_unstemmed | CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L) |
| title_short | CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L) |
| title_sort | crispr cas9 gene editing of a sox9 reporter human ipsc line to produce two trpv4 patient heterozygous missense mutant ipsc lines mcrii001 a 3 trpv4 p f273l and mcrii001 a 4 trpv4 p p799l |
| url | http://www.sciencedirect.com/science/article/pii/S1873506120302439 |
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