| 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
|
| リソース情報 |
| ヒト・動物細胞 |
293T(RCB2202) |
