Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking
The most highly studied endocytic pathway, clathrin-dependent endocytosis, mediates a wide range of fundamental processes including nutrient internalization, receptor recycling, and signal transduction. In order to model tissue specific and developmental aspects of this process, CRISPR/Cas9 genomic...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-12-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506118302423 |
| _version_ | 1818213282974007296 |
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| author | Ruthellen H. Anderson Jason G. Kerkvliet Jaelin J. Otta Alan D. Ross Patricia C. Leiferman Adam D. Hoppe Kevin R. Francis |
| author_facet | Ruthellen H. Anderson Jason G. Kerkvliet Jaelin J. Otta Alan D. Ross Patricia C. Leiferman Adam D. Hoppe Kevin R. Francis |
| author_sort | Ruthellen H. Anderson |
| collection | DOAJ |
| description | The most highly studied endocytic pathway, clathrin-dependent endocytosis, mediates a wide range of fundamental processes including nutrient internalization, receptor recycling, and signal transduction. In order to model tissue specific and developmental aspects of this process, CRISPR/Cas9 genomic editing was utilized to fluorescently label the C-terminus of clathrin light chain A (CLTA) within the phenotypically normal, parental CRMi001-A human induced pluripotent stem cell line. Successfully edited cells were isolated by fluorescently activated cell sorting, remained karyotypically normal, and maintained their differentiation potential. This cell line facilitates imaging of endogenous clathrin trafficking within varied cell types. |
| first_indexed | 2024-12-12T06:01:49Z |
| format | Article |
| id | doaj.art-3e5c5a930c714f57b390c58bf4e1ec9f |
| institution | Directory Open Access Journal |
| issn | 1873-5061 |
| language | English |
| last_indexed | 2024-12-12T06:01:49Z |
| publishDate | 2018-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-3e5c5a930c714f57b390c58bf4e1ec9f2022-12-22T00:35:23ZengElsevierStem Cell Research1873-50612018-12-01339599Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin traffickingRuthellen H. Anderson0Jason G. Kerkvliet1Jaelin J. Otta2Alan D. Ross3Patricia C. Leiferman4Adam D. Hoppe5Kevin R. Francis6Sanford School of Medicine, University of South Dakota, Sioux Falls, SD, USA; Cellular Therapies and Stem Cell Biology Group, Sanford Research, Sioux Falls, SD, USADepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD, USA; BioSystems Networks and Translational Research Center, Brookings, SD, USASanford School of Medicine, University of South Dakota, Sioux Falls, SD, USASanford Medical Genetics Laboratory, Cytogenetics Division, Sanford Hospital, Sioux Falls, SD, USASanford School of Medicine, University of South Dakota, Sioux Falls, SD, USA; Sanford Medical Genetics Laboratory, Cytogenetics Division, Sanford Hospital, Sioux Falls, SD, USADepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD, USA; BioSystems Networks and Translational Research Center, Brookings, SD, USASanford School of Medicine, University of South Dakota, Sioux Falls, SD, USA; Cellular Therapies and Stem Cell Biology Group, Sanford Research, Sioux Falls, SD, USA; Corresponding author at: Sanford School of Medicine, University of South Dakota, Sioux Falls, SD, USA.The most highly studied endocytic pathway, clathrin-dependent endocytosis, mediates a wide range of fundamental processes including nutrient internalization, receptor recycling, and signal transduction. In order to model tissue specific and developmental aspects of this process, CRISPR/Cas9 genomic editing was utilized to fluorescently label the C-terminus of clathrin light chain A (CLTA) within the phenotypically normal, parental CRMi001-A human induced pluripotent stem cell line. Successfully edited cells were isolated by fluorescently activated cell sorting, remained karyotypically normal, and maintained their differentiation potential. This cell line facilitates imaging of endogenous clathrin trafficking within varied cell types.http://www.sciencedirect.com/science/article/pii/S1873506118302423 |
| spellingShingle | Ruthellen H. Anderson Jason G. Kerkvliet Jaelin J. Otta Alan D. Ross Patricia C. Leiferman Adam D. Hoppe Kevin R. Francis Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking Stem Cell Research |
| title | Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking |
| title_full | Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking |
| title_fullStr | Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking |
| title_full_unstemmed | Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking |
| title_short | Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking |
| title_sort | generation of a clta reporter human induced pluripotent stem cell line crmi001 a 1 using the crispr cas9 system to monitor endogenous clathrin trafficking |
| url | http://www.sciencedirect.com/science/article/pii/S1873506118302423 |
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