Generation of a rod-specific NRL reporter line in human pluripotent stem cells

Abstract Reporter lines generated in human pluripotent stem cells can be highly useful for the analysis of specific cell types and lineages in live cultures. We created the first human rod reporter line using CRISPR/Cas9 genome editing to replace one allele of the Neural Retina Leucine zipper (NRL)...

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Autores principales: M. Joseph Phillips, Elizabeth E. Capowski, Andrew Petersen, Alex D. Jansen, Katherine Barlow, Kimberly L. Edwards, David M. Gamm
Formato: article
Lenguaje:EN
Publicado: Nature Portfolio 2018
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Acceso en línea:https://doaj.org/article/bc6bee677a18423c99acf52dbb951ec4
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spelling oai:doaj.org-article:bc6bee677a18423c99acf52dbb951ec42021-12-02T15:08:55ZGeneration of a rod-specific NRL reporter line in human pluripotent stem cells10.1038/s41598-018-20813-32045-2322https://doaj.org/article/bc6bee677a18423c99acf52dbb951ec42018-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-20813-3https://doaj.org/toc/2045-2322Abstract Reporter lines generated in human pluripotent stem cells can be highly useful for the analysis of specific cell types and lineages in live cultures. We created the first human rod reporter line using CRISPR/Cas9 genome editing to replace one allele of the Neural Retina Leucine zipper (NRL) gene with an eGFP transgene in the WA09 human embryonic stem cell (hESC) line. After confirming successful targeting, three-dimensional optic vesicle structures were produced to examine reporter specificity and to track rod differentiation in culture. The NRL +/eGFP hESC line robustly and exclusively labeled the entirety of rods throughout differentiation, eventually revealing highly mature structural features. This line provides a valuable tool for studying human rod development and disease and testing therapeutic strategies for retinitis pigmentosa.M. Joseph PhillipsElizabeth E. CapowskiAndrew PetersenAlex D. JansenKatherine BarlowKimberly L. EdwardsDavid M. GammNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
M. Joseph Phillips
Elizabeth E. Capowski
Andrew Petersen
Alex D. Jansen
Katherine Barlow
Kimberly L. Edwards
David M. Gamm
Generation of a rod-specific NRL reporter line in human pluripotent stem cells
description Abstract Reporter lines generated in human pluripotent stem cells can be highly useful for the analysis of specific cell types and lineages in live cultures. We created the first human rod reporter line using CRISPR/Cas9 genome editing to replace one allele of the Neural Retina Leucine zipper (NRL) gene with an eGFP transgene in the WA09 human embryonic stem cell (hESC) line. After confirming successful targeting, three-dimensional optic vesicle structures were produced to examine reporter specificity and to track rod differentiation in culture. The NRL +/eGFP hESC line robustly and exclusively labeled the entirety of rods throughout differentiation, eventually revealing highly mature structural features. This line provides a valuable tool for studying human rod development and disease and testing therapeutic strategies for retinitis pigmentosa.
format article
author M. Joseph Phillips
Elizabeth E. Capowski
Andrew Petersen
Alex D. Jansen
Katherine Barlow
Kimberly L. Edwards
David M. Gamm
author_facet M. Joseph Phillips
Elizabeth E. Capowski
Andrew Petersen
Alex D. Jansen
Katherine Barlow
Kimberly L. Edwards
David M. Gamm
author_sort M. Joseph Phillips
title Generation of a rod-specific NRL reporter line in human pluripotent stem cells
title_short Generation of a rod-specific NRL reporter line in human pluripotent stem cells
title_full Generation of a rod-specific NRL reporter line in human pluripotent stem cells
title_fullStr Generation of a rod-specific NRL reporter line in human pluripotent stem cells
title_full_unstemmed Generation of a rod-specific NRL reporter line in human pluripotent stem cells
title_sort generation of a rod-specific nrl reporter line in human pluripotent stem cells
publisher Nature Portfolio
publishDate 2018
url https://doaj.org/article/bc6bee677a18423c99acf52dbb951ec4
work_keys_str_mv AT mjosephphillips generationofarodspecificnrlreporterlineinhumanpluripotentstemcells
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AT alexdjansen generationofarodspecificnrlreporterlineinhumanpluripotentstemcells
AT katherinebarlow generationofarodspecificnrlreporterlineinhumanpluripotentstemcells
AT kimberlyledwards generationofarodspecificnrlreporterlineinhumanpluripotentstemcells
AT davidmgamm generationofarodspecificnrlreporterlineinhumanpluripotentstemcells
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