Project description:Purpose: We aimed to identify the targets of the RNA binding protein TruB1 in HEK293FT cells. Methods: Total RNA from endogenous-Flag KI cells were treated with UV light to crosslink RNA and proteins, then RNA-protein complexes were pulled down with anti-Flag. RNA was extracted and used to make cDNS libraries that were then sequenced by MiSeq 150bp single-end read (Sample1). Results: Reads were trimmed to remove any adaptor sequence and barcodes before mapping reads to genome hg38 using Bowtie2. Conclusions: We identified TruB1 binding sites in tRNAs and noncodingRNAs.
Project description:In order to identify YBX1 binding sites on tRNA fragments, we performed small-RNA HITS-CLIP on endogenous YBX1 We used a previously published method to perform HITS-CLIP on endogenous YBX1 (Chi SW, et al. 2009, Nature 460:479)
Project description:In order to identify YBX1 binding sites on endogenous RNA, we performed HITS-CLIP on endogenous YBX1 We used a previously published method to perform HITS-CLIP on endogenous YBX1 (Licatalosi D, et al. 2008, Nature 456:464-U22)
Project description:In order to identify TARBP2 binding sites on endogenous RNA, we performed HITS-CLIP on a myc-tagged TARBP2 expressing cell-line (transient transfection) Cells were transfected with tagged TARBP2 vector (Origene) and 48-hr post-transfection, they were subjected to the HITS-CLIP procedure (Licatalosi D, et al. 2008, Nature 456:464-U22)