Multiplexed transcriptome discovery of RNA-binding protein binding sites by antibody-barcode eCLIP.
Nat Methods
; 20(1): 65-69, 2023 01.
Article
em En
| MEDLINE
| ID: mdl-36550273
ABSTRACT
Ultraviolet crosslinking and immunoprecipitation (CLIP) methodologies enable the identification of RNA binding sites of RNA-binding proteins (RBPs). Despite improvements in the library preparation of RNA fragments, the enhanced CLIP (eCLIP) protocol requires 4 days of hands-on time and lacks the ability to process several RBPs in parallel. We present a new method termed antibody-barcode eCLIP that utilizes DNA-barcoded antibodies and proximity ligation of the DNA oligonucleotides to RBP-protected RNA fragments to interrogate several RBPs simultaneously. We observe performance comparable with that of eCLIP with the advantage of dramatically increased scaling while maintaining the same material requirement of a single eCLIP experiment.
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Base de dados:
MEDLINE
Assunto principal:
RNA
/
Transcriptoma
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article