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...

Descripción completa

Detalles Bibliográficos
Autores principales: Anderson, Ruthellen H., Kerkvliet, Jason G., Otta, Jaelin J., Ross, Alan D., Leiferman, Patricia C., Hoppe, Adam D., Francis, Kevin R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2018
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6383648/
https://www.ncbi.nlm.nih.gov/pubmed/30340091
http://dx.doi.org/10.1016/j.scr.2018.10.001
_version_ 1783396876725256192
author Anderson, Ruthellen H.
Kerkvliet, Jason G.
Otta, Jaelin J.
Ross, Alan D.
Leiferman, Patricia C.
Hoppe, Adam D.
Francis, Kevin R.
author_facet Anderson, Ruthellen H.
Kerkvliet, Jason G.
Otta, Jaelin J.
Ross, Alan D.
Leiferman, Patricia C.
Hoppe, Adam D.
Francis, Kevin R.
author_sort Anderson, Ruthellen H.
collection PubMed
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.
format Online
Article
Text
id pubmed-6383648
institution National Center for Biotechnology Information
language English
publishDate 2018
record_format MEDLINE/PubMed
spelling pubmed-63836482019-02-21 Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking Anderson, Ruthellen H. Kerkvliet, Jason G. Otta, Jaelin J. Ross, Alan D. Leiferman, Patricia C. Hoppe, Adam D. Francis, Kevin R. Stem Cell Res Article 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. 2018-10-03 2018-12 /pmc/articles/PMC6383648/ /pubmed/30340091 http://dx.doi.org/10.1016/j.scr.2018.10.001 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/BY-NC-ND/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) ).
spellingShingle Article
Anderson, Ruthellen H.
Kerkvliet, Jason G.
Otta, Jaelin J.
Ross, Alan D.
Leiferman, Patricia C.
Hoppe, Adam D.
Francis, Kevin R.
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 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6383648/
https://www.ncbi.nlm.nih.gov/pubmed/30340091
http://dx.doi.org/10.1016/j.scr.2018.10.001
work_keys_str_mv AT andersonruthellenh generationofacltareporterhumaninducedpluripotentstemcelllinecrmi001a1usingthecrisprcas9systemtomonitorendogenousclathrintrafficking
AT kerkvlietjasong generationofacltareporterhumaninducedpluripotentstemcelllinecrmi001a1usingthecrisprcas9systemtomonitorendogenousclathrintrafficking
AT ottajaelinj generationofacltareporterhumaninducedpluripotentstemcelllinecrmi001a1usingthecrisprcas9systemtomonitorendogenousclathrintrafficking
AT rossaland generationofacltareporterhumaninducedpluripotentstemcelllinecrmi001a1usingthecrisprcas9systemtomonitorendogenousclathrintrafficking
AT leifermanpatriciac generationofacltareporterhumaninducedpluripotentstemcelllinecrmi001a1usingthecrisprcas9systemtomonitorendogenousclathrintrafficking
AT hoppeadamd generationofacltareporterhumaninducedpluripotentstemcelllinecrmi001a1usingthecrisprcas9systemtomonitorendogenousclathrintrafficking
AT franciskevinr generationofacltareporterhumaninducedpluripotentstemcelllinecrmi001a1usingthecrisprcas9systemtomonitorendogenousclathrintrafficking

Ejemplares similares