論文 - 詳細
| RRC ID | 83531 |
|---|---|
| 著者 | Kunitake K, Mizuno T, Hattori K, Oneyama C, Kamiya M, Ota S, Urano Y, Kojima R. |
| タイトル | Barcoding of small extracellular vesicles with CRISPR-gRNA enables comprehensive, subpopulation-specific analysis of their biogenesis and release regulators. |
| ジャーナル | Nat Commun |
| Abstract |
Small extracellular vesicles (sEVs) are important intercellular information transmitters in various biological contexts, but their release processes remain poorly understood. Herein, we describe a high-throughput assay platform, CRISPR-assisted individually barcoded sEV-based release regulator (CIBER) screening, for identifying key players in sEV release. CIBER screening employs sEVs barcoded with CRISPR-gRNA through the interaction of gRNA and dead Cas9 fused with an sEV marker. Barcode quantification enables the estimation of the sEV amount released from each cell in a massively parallel manner. Barcoding sEVs with different sEV markers in a CRISPR pooled-screening format allows genome-wide exploration of sEV release regulators in a subpopulation-specific manner, successfully identifying previously unknown sEV release regulators and uncovering the exosomal/ectosomal nature of CD63+/CD9+ sEVs, respectively, as well as the synchronization of CD9+ sEV release with the cell cycle. CIBER should be a valuable tool for detailed studies on the biogenesis, release, and heterogeneity of sEVs. |
| 巻・号 | 15(1) |
| ページ | 9777 |
| 公開日 | 2024-11-19 |
| DOI | 10.1038/s41467-024-53736-x |
| PII | 10.1038/s41467-024-53736-x |
| PMID | 39562573 |
| PMC | PMC11577021 |
| MeSH | CRISPR-Cas Systems* Clustered Regularly Interspaced Short Palindromic Repeats / genetics Exosomes / genetics Exosomes / metabolism Extracellular Vesicles* / genetics Extracellular Vesicles* / metabolism HEK293 Cells Humans RNA, Guide, CRISPR-Cas Systems* / genetics RNA, Guide, CRISPR-Cas Systems* / metabolism Tetraspanin 29* / genetics Tetraspanin 29* / metabolism Tetraspanin 30* / genetics Tetraspanin 30* / metabolism |
| IF | 12.121 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 40 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | 293T(RCB2202) |