RRC ID 21530
著者 Shin JW, Suzuki T, Ninomiya N, Kishima M, Hasegawa Y, Kubosaki A, Yabukami H, Hayashizaki Y, Suzuki H.
タイトル Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming.
ジャーナル Nucleic Acids Res
Abstract 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.
巻・号 40(21)
ページ e165
公開日 2012-11-1
DOI 10.1093/nar/gks732
PII gks732
PMID 22879381
PMC PMC3505982
MeSH Cell Transdifferentiation / genetics* Cells, Cultured Fibroblasts / metabolism Gene Expression Humans Lentivirus / genetics Monocytes / metabolism Proto-Oncogene Proteins / genetics Proto-Oncogene Proteins / metabolism Single-Cell Analysis / methods* Trans-Activators / genetics Trans-Activators / metabolism Transcription, Genetic*
IF 11.502
引用数 7
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
遺伝子材料 Genome Network Project Gateway Entry Clone W01A062B19 (HGE024843) W01A003N09 (HGE001521) W01A126A07 (HGE050407) W01A125A03 (HGE050003) W01A004O18 (HGE001954) W01A006J20 (HGE002636) W01A044B11 (HGE017635) W01A003I20 (HGE001412) W01A002L03 (HGE001067)