| RRC ID |
54060
|
| 著者 |
Nishioka K, Miyazaki H, Soejima H.
|
| タイトル |
Unbiased shRNA screening, using a combination of FACS and high-throughput sequencing, enables identification of novel modifiers of Polycomb silencing.
|
| ジャーナル |
Sci Rep
|
| Abstract |
Polycomb silencing is an important and rapidly growing field that is relevant to a broad range of aspects of human health, including cancer and stem cell biology. To date, the regulatory mechanisms for the fine-tuning of Polycomb silencing remain unclear, but it is likely that there is a series of unidentified factors that functionally modify or balance the silencing. However, a practical gene screening strategy for identifying such factors has not yet been developed. The failure of screening strategies used thus far is probably due to the effect of the loss-of-function phenotypes of these factors on cell cycle progression. Here, by applying fluorescence-activated cell sorter (FACS) and high-throughput sequencing (HTS) technology in a large-scale lentivirus-mediated shRNA screening, we obtained a consecutive dataset from all shRNAs tested, which highlighted a substantial number of genes that may control Polycomb silencing. We consider that this unbiased strategy can readily be applied to a wide range of studies to uncover novel regulatory layers for expression of genes of interest.
|
| 巻・号 |
8(1)
|
| ページ |
12128
|
| 公開日 |
2018-8-14
|
| DOI |
10.1038/s41598-018-30649-6
|
| PII |
10.1038/s41598-018-30649-6
|
| PMID |
30108332
|
| PMC |
PMC6092423
|
| MeSH |
Animals
Cell Line, Tumor
Cell Separation / methods
Datasets as Topic
Flow Cytometry / methods
Genes, Reporter / genetics
Genetic Vectors / genetics
High-Throughput Nucleotide Sequencing / methods
High-Throughput Screening Assays / methods*
Lentivirus / genetics
Mice
Mouse Embryonic Stem Cells
Polycomb-Group Proteins / genetics*
RNA Interference
RNA, Small Interfering / metabolism*
|
| IF |
3.998
|
| 引用数 |
4
|
| リソース情報 |
| ヒト・動物細胞 |
F9(RCB1555) |
