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MARIS: Method for Analyzing RNA following Intracellular Sorting

Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high...

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Detalles Bibliográficos
Autores principales: Hrvatin, Siniša, Deng, Francis, O'Donnell, Charles W., Gifford, David K., Melton, Douglas A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3940959/
https://www.ncbi.nlm.nih.gov/pubmed/24594682
http://dx.doi.org/10.1371/journal.pone.0089459
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author Hrvatin, Siniša
Deng, Francis
O'Donnell, Charles W.
Gifford, David K.
Melton, Douglas A.
author_facet Hrvatin, Siniša
Deng, Francis
O'Donnell, Charles W.
Gifford, David K.
Melton, Douglas A.
author_sort Hrvatin, Siniša
collection PubMed
description Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human β cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines.
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spelling pubmed-39409592014-03-06 MARIS: Method for Analyzing RNA following Intracellular Sorting Hrvatin, Siniša Deng, Francis O'Donnell, Charles W. Gifford, David K. Melton, Douglas A. PLoS One Research Article Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human β cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines. Public Library of Science 2014-03-03 /pmc/articles/PMC3940959/ /pubmed/24594682 http://dx.doi.org/10.1371/journal.pone.0089459 Text en © 2014 Hrvatin et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Hrvatin, Siniša
Deng, Francis
O'Donnell, Charles W.
Gifford, David K.
Melton, Douglas A.
MARIS: Method for Analyzing RNA following Intracellular Sorting
title MARIS: Method for Analyzing RNA following Intracellular Sorting
title_full MARIS: Method for Analyzing RNA following Intracellular Sorting
title_fullStr MARIS: Method for Analyzing RNA following Intracellular Sorting
title_full_unstemmed MARIS: Method for Analyzing RNA following Intracellular Sorting
title_short MARIS: Method for Analyzing RNA following Intracellular Sorting
title_sort maris: method for analyzing rna following intracellular sorting
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3940959/
https://www.ncbi.nlm.nih.gov/pubmed/24594682
http://dx.doi.org/10.1371/journal.pone.0089459
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