GoldCLIP: Gel-omitted Ligation-dependent CLIP / 基因组蛋白质组与生物信息学报·英文版
Genomics, Proteomics & Bioinformatics
;
(4): 136-143, 2018.
Artículo
en Inglés
| WPRIM
| ID: wpr-773000
ABSTRACT
Protein-RNA interaction networks are essential to understand gene regulation control. Identifying binding sites of RNA-binding proteins (RBPs) by the UV-crosslinking and immunoprecipitation (CLIP) represents one of the most powerful methods to map protein-RNA interactions in vivo. However, the traditional CLIP protocol is technically challenging, which requires radioactive labeling and suffers from material loss during PAGE-membrane transfer procedures. Here we introduce a super-efficient CLIP method (GoldCLIP) that omits all gel purification steps. This nonisotopic method allows us to perform highly reproducible CLIP experiments with polypyrimidine tract-binding protein (PTB), a classical RBP in human cell lines. In principle, our method guarantees sequencing library constructions, providing the protein of interest can be successfully crosslinked to RNAs in living cells. GoldCLIP is readily applicable to diverse proteins to uncover their endogenous RNA targets.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Sitios de Unión
/
ARN
/
Línea Celular
/
Proteínas de Unión al ARN
/
Proteína de Unión al Tracto de Polipirimidina
/
Ribonucleoproteínas Nucleares Heterogéneas
/
Inmunoprecipitación
/
Metabolismo
/
Métodos
Tipo de estudio:
Estudio pronóstico
Límite:
Humanos
Idioma:
Inglés
Revista:
Genomics, Proteomics & Bioinformatics
Año:
2018
Tipo del documento:
Artículo
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