| RRC ID |
89439
|
| 著者 |
Hu LF, Xie G, Wu YX, Li YX, Wan ZL, Mi L, Wang JZ, Wang Y.
|
| タイトル |
Longitudinal monitoring of cytoplasmic RBP-RNA interactions and transcriptome in living cells by engineered protein nanocages.
|
| ジャーナル |
Mol Cell
|
| Abstract |
Most existing RNA sequencing methods rely on cell lysis or fixation, limiting their use in longitudinal studies of the same cell population. Here, we introduce POND-seq (protein nanocage-empowered non-destructive sequencing), a strategy that employs secretory protein nanocages fused to RNA-binding proteins (RBPs) to recover RBP-associated RNAs from living cells. POND-seq robustly identifies RNA targets of cytoplasmic RBPs across multiple cell types and enables longitudinal tracking of dynamic changes in RBP-associated RNA profiles under stress conditions. Fusion to poly(A)-binding protein C (PABPC1) further allows monitoring of transcriptomic responses and selectively profiles cell-type-specific transcriptomes from mixed populations without cell dissociation and sorting. Additionally, POND-seq supports functional interrogation of RBP domains and residues involved in RNA association and enables scalable analysis of RBP variants, as demonstrated by a systematic assessment of disease-associated fragile X messenger ribonucleoprotein 1 (FMR1) mutations. Together, POND-seq provides a versatile and scalable platform for non-destructive and longitudinal analysis of cytoplasmic transcriptomes and RBP-associated RNAs.
|
| 公開日 |
2026-6-25
|
| DOI |
10.1016/j.molcel.2026.06.007
|
| PII |
S1097-2765(26)00378-3
|
| PMID |
42349405
|
| リソース情報 |
| ヒト・動物細胞 |
HuH-7(RCB1366) |
