Reference - Detail
| RRC ID | 21530 |
|---|---|
| Author | Shin JW, Suzuki T, Ninomiya N, Kishima M, Hasegawa Y, Kubosaki A, Yabukami H, Hayashizaki Y, Suzuki H. |
| Title | Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming. |
| Journal | 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. |
| Volume | 40(21) |
| Pages | e165 |
| Published | 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 |
| Times Cited | 7 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 12 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | Genome Network Project Gateway Entry Clone W01A062B19 (HGE024843) W01A003N09 (HGE001521) W01A126A07 (HGE050407) W01A125A03 (HGE050003) W01A004O18 (HGE001954) W01A006J20 (HGE002636) W01A044B11 (HGE017635) W01A003I20 (HGE001412) W01A002L03 (HGE001067) |
| Human and Animal Cells | Human dermal fibroblasts |