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Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming
Combinatorial interactions of transcription modulators are critical to regulate cell-specific expression and to drive direct cell reprogramming (e.g. trans-differentiation). However, the identification of key transcription modulators from myriad of candidate genes is laborious and time consuming. To...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3505982/ https://www.ncbi.nlm.nih.gov/pubmed/22879381 http://dx.doi.org/10.1093/nar/gks732 |
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author | Shin, Jay W. Suzuki, Takahiro Ninomiya, Noriko Kishima, Mami Hasegawa, Yuki Kubosaki, Atsutaka Yabukami, Haruka Hayashizaki, Yoshihide Suzuki, Harukazu |
author_facet | Shin, Jay W. Suzuki, Takahiro Ninomiya, Noriko Kishima, Mami Hasegawa, Yuki Kubosaki, Atsutaka Yabukami, Haruka Hayashizaki, Yoshihide Suzuki, Harukazu |
author_sort | Shin, Jay W. |
collection | PubMed |
description | Combinatorial interactions of transcription modulators are critical to regulate cell-specific expression and to drive direct cell reprogramming (e.g. trans-differentiation). However, the identification of key transcription modulators from myriad of candidate genes is laborious and time consuming. To rapidly identify key regulatory factors involved in direct cell reprogramming, we established a multiplex single-cell screening system using a fibroblast-to-monocyte transition model. The system implements a single-cell ‘shotgun-transduction’ strategy followed by nested-single-cell-polymerase chain reaction (Nesc-PCR) gene expression analysis. To demonstrate this, we simultaneously transduced 18 monocyte-enriched transcription modulators in fibroblasts followed by selection of single cells expressing monocyte-specific CD14 and HLA-DR cell-surface markers from a heterogeneous population. Highly multiplex Nesc-PCR expression analysis revealed a variety of gene combinations with a significant enrichment of SPI1 (86/86) and a novel transcriptional modulator, HCLS1 (76/86), in the CD14(+)/HLA-DR(+) single cells. We could further demonstrate the synergistic role of HCLS1 in regulating monocyte-specific gene expressions and phagocytosis in dermal fibroblasts in the presence of SPI1. This study establishes a platform for a multiplex single-cell screening of combinatorial transcription modulators to drive any direct cell reprogramming. |
format | Online Article Text |
id | pubmed-3505982 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-35059822012-11-26 Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming Shin, Jay W. Suzuki, Takahiro Ninomiya, Noriko Kishima, Mami Hasegawa, Yuki Kubosaki, Atsutaka Yabukami, Haruka Hayashizaki, Yoshihide Suzuki, Harukazu Nucleic Acids Res Methods Online Combinatorial interactions of transcription modulators are critical to regulate cell-specific expression and to drive direct cell reprogramming (e.g. trans-differentiation). However, the identification of key transcription modulators from myriad of candidate genes is laborious and time consuming. To rapidly identify key regulatory factors involved in direct cell reprogramming, we established a multiplex single-cell screening system using a fibroblast-to-monocyte transition model. The system implements a single-cell ‘shotgun-transduction’ strategy followed by nested-single-cell-polymerase chain reaction (Nesc-PCR) gene expression analysis. To demonstrate this, we simultaneously transduced 18 monocyte-enriched transcription modulators in fibroblasts followed by selection of single cells expressing monocyte-specific CD14 and HLA-DR cell-surface markers from a heterogeneous population. Highly multiplex Nesc-PCR expression analysis revealed a variety of gene combinations with a significant enrichment of SPI1 (86/86) and a novel transcriptional modulator, HCLS1 (76/86), in the CD14(+)/HLA-DR(+) single cells. We could further demonstrate the synergistic role of HCLS1 in regulating monocyte-specific gene expressions and phagocytosis in dermal fibroblasts in the presence of SPI1. This study establishes a platform for a multiplex single-cell screening of combinatorial transcription modulators to drive any direct cell reprogramming. Oxford University Press 2012-11 2012-08-08 /pmc/articles/PMC3505982/ /pubmed/22879381 http://dx.doi.org/10.1093/nar/gks732 Text en © The Author(s) 2012. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methods Online Shin, Jay W. Suzuki, Takahiro Ninomiya, Noriko Kishima, Mami Hasegawa, Yuki Kubosaki, Atsutaka Yabukami, Haruka Hayashizaki, Yoshihide Suzuki, Harukazu Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming |
title | Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming |
title_full | Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming |
title_fullStr | Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming |
title_full_unstemmed | Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming |
title_short | Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming |
title_sort | establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3505982/ https://www.ncbi.nlm.nih.gov/pubmed/22879381 http://dx.doi.org/10.1093/nar/gks732 |
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