MS2-RIP Assay for Identifying the Interaction Partners of lncRNA DANCR in Tumor Cells
- VernacularTitle:构建MS2-RIP方法用于鉴定lncRNA DANCR在肿瘤细胞中的相互作用分子
- Author:
Meng-shi WU
1
;
Min-min XIONG
1
;
Dan PENG
1
;
Xue HAN
1
;
Xiao-min ZHONG
1
Author Information
1. Center for Stem Cell Biology and Tissue Engineering，Zhongshan School of Medicine，Sun Yat-Sen University，Guangzhou 510080，China
- Publication Type:Journal Article
- Keywords:
MS2;
RNA-binding protein immunoprecipitation（RIP）;
long non-coding RNA（lncRNA）;
DANCR
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2021;42(1):24-32
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the functional mechanism of lncRNA DANCR in tumor cells， a MS2-RIP method was designed and conducted to identify the molecules that interact with DANCR. MethodsThe specific binding of MS2 bacteriophage capsid protein and MS2 binding site （MS2 BS）， a 19-base RNA stem-loop structure located at the 5′ terminus of the MS2 bacteriophage replicase gene， was applied to construct a plasmid for tandemly overexpressing DANCR and MS2 BS. DANCR was enriched and its associated molecules were further identified and analyzed. ResultsThe plasmid for tandemly overexpressing DANCR and MS2 BS was successfully constructed for the MS2-RIP experiment， which could significantly enrich DANCR and DANCR-binding protein EZH2 as a positive control. ConclusionBased on the high affinity binding between MS2 protein and MS2 BS， the MS2-RIP method was established for lncRNA DANCR， which could effectively capture DANCR as well as its associated molecules， providing a new technology for studying the functional mechanism of DANCR.
