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MaTAR25 RNA Antisense Pulldown and Mass Spectrometry

ABSTRACT: It has been suggested that lncRNAs can interact with transcriptional regulators/co-factors to form ribonucleoprotein (RNP) complexes to regulate the expression of downstream genes in the cell nucleus. To identify potential interacting proteins of MaTAR25, we used two different paired sets of biotin-labeled antisense oligonucleotides targeting MaTAR25 for native RNA antisense oligonucleotide pull-down (RAP) in 4T1 cells followed by qRT-PCR to assess pull-down efficiency. Samples were also eluted from beads for mass spectrometry isobaric tags for relative and absolute quantitative (MS-iTRAQ) analysis to identify proteins that bind to MaTAR25, and PPIB as the corresponding control. We ranked the candidate interactors based on detectable peptides above background in both pair sets of oligonucleotide pull-downs, and selected candidates with at least 2-fold enrichment compared to corresponding PPIB oligo pull-down.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: Kung-Chi Chang

LAB HEAD: David L. Spector

PROVIDER: PXD017398 | Pride | 2020-12-23

REPOSITORIES: Pride

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			Action	DRS
	F013445.dat	Other
	MaTAR25_FX_01_181003131922_ppib.raw	Raw
	MaTAR25_FX_02M25oligo10.raw	Raw
	MaTAR25_FX_03M25oligo7_8.raw	Raw
	MaTAR25_FX_04M25oligo6_10.raw	Raw

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