采用MS2 pulldown和质谱分析法检测BT549细胞中的LncRNASNHG15结合蛋白

doi:10.3969/j.issn.1004-616x.2024.05.010

Abstract

Abstract: OBJECTIVE： To identify proteins which interacted with long noncoding RNA SNHG15 in breast cancer cells. METHODS： BT549 cells stably overexpressing SNHG15 were established by infection with lentivirus expressing MS2bs-tagged SHNG15. MS2 pulldown and Mass Spectrometry were used to identify proteins associated with SNHG15. Using DDX5 (DEAD box helicase 5) as an example，interactions of SHNG15 with identified proteins were further verified by RNA immunoprecipitation(RIP)and RT-qPCR. RESULTS： In total，386 proteins were identified which interacted with SNHG15. DDX5 was truly associated with SNHG15. CONCLUSION： These data indicated an effective method to purify RNA-associated proteins and provided new information for the further study of the function of SNHG15.

Key words: long noncoding RNA, MS2 pulldown, SNHG15, RNA immunoprecipitation, RNA binding protein

CLC Number:

Q291

CHEN Shaoying, PENG Pei, ZHOU Yanchun, GU Wei. Identification of LncRNA SNHG15 binding proteins in BT549 cells using MS2 pulldown and mass spectrometry[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(5): 395-398.

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Identification of LncRNA SNHG15 binding proteins in BT549 cells using MS2 pulldown and mass spectrometry

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