| 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
|
| 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 |
