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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Main Authors:	M. Joseph Phillips, Elizabeth E. Capowski, Andrew Petersen, Alex D. Jansen, Katherine Barlow, Kimberly L. Edwards, David M. Gamm
Format:	article
Language:	EN
Published:	Nature Portfolio 2018
Subjects:	Medicine R Science Q
Online Access:	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
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_version_	1718387962909032448

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

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