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.
Artigo
em Inglês
| 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:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Sítios de Ligação
/
RNA
/
Linhagem Celular
/
Proteínas de Ligação a RNA
/
Proteína de Ligação a Regiões Ricas em Polipirimidinas
/
Ribonucleoproteínas Nucleares Heterogêneas
/
Imunoprecipitação
/
Metabolismo
/
Métodos
Tipo de estudo:
Estudo prognóstico
Limite:
Humanos
Idioma:
Inglês
Revista:
Genomics, Proteomics & Bioinformatics
Ano de publicação:
2018
Tipo de documento:
Artigo
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