Biochemical analysis of long non-coding RNA-containing ribonucleoprotein complexes.
Long non-coding RNAs (lncRNAs), once relegated to junk products of the genome, are becoming better appreciated for the myriad functions they play in cellular processes. It is clear that for most of the cases studied, lncRNAs carry out their functions at least in part through interactions with proteins. Here we present two complementary biochemical methods for the analysis of lncRNA-containing ribonucleoprotein complexes, hereafter referred to as RNPs. The first strategy offers users the ability to purify RNPs based on a protein component and to analyze the spectrum of lncRNAs, other proteins, and, if present, other types of RNAs that are bound to it. The second makes use of a bacteriophage MS2 binding-site affinity-handle grafted onto an lncRNA of interest to investigate the proteins and RNAs that co-purify with the tagged RNA.
Department of Biochemistry and Biophysics, School of Medicine and Dentistry, Center for RNA Biology, University of Rochester, Rochester, NY 14642, USA.
Chenguang
C
Gong
Maximilian Wei-Lin
MW
Popp
Lynne E
LE
Maquat
D001665:Binding Sites; D017909:Levivirus / Q000235:genetics; D062085:RNA, Long Noncoding / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D012261:Ribonucleoproteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification
D062085:RNA, Long Noncoding; D012261:Ribonucleoproteins