Project description:The RNA- and DNA-binding protein YBX1 is involved in various cellular processes. We here focused on investigating its role in posttranscriptional gene regulation in medulloblastoma, in which CRISPR screens have shown its essentiality for cellular survival. We found that YBX1 knockdown leads to broad upregulation of genes related to neuroinflammation, and directly regulates mRNA stability of various mRNA targets. More specifically, target CBX5, also called heterochromatin-protein 1 alpha (HP1-alpha), is downregulated upon YBX1 KD. It associates strongly with heterochromatin, and upon downregulation allows for upregulation of heterochromatin repressed genes related to inflammation.
Project description:The RNA- and DNA-binding protein YBX1 is involved in various cellular processes. We here focused on investigating its role in posttranscriptional gene regulation in medulloblastoma, in which CRISPR screens have shown its essentiality for cellular survival. We found that YBX1 knockdown leads to broad upregulation of genes related to neuroinflammation, and directly regulates mRNA stability of various mRNA targets. More specifically, target CBX5, also called heterochromatin-protein 1 alpha (HP1-alpha), is downregulated upon YBX1 KD. It associates strongly with heterochromatin, and upon downregulation allows for upregulation of heterochromatin repressed genes related to inflammation.
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 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:YBX1 is a multifunctional protein involved in the control of transcription and translation. We identified YBX1 as an target of MEK/ERK signaling in colorectal cancer cell lines. We performed a ChIP-chip analysis of HCT116 cells to identify new potential target genes of YBX1. Comparison of input DNA fragments with fragments coprecipitated with YBX1 in HCT116 cells.
Project description:In order to identify YBX1-dependent targets that are modulated under hypoxic conditions, we used control and YBX1 knockdown cells grown under normoxia and hypoxia to profile gene expression levels. Control and YBX1-knockdown cells were grown and profiled under hypoxia and normoxia to identify YBX1-dependent hypoxia-induced target transcripts.
Project description:BRD4 is amplified and/or up-regiulated in a subset of ovarian cancer which correlates with a poor survival siRNA sensitization screens in the presence of CHK1 inhibitor, LY2606368, identify BRD4 as a therapeutic target in ovarian cancer. BRD4 suppression by either siRNA or using JQ1 increases CBX5 expression
Project description:In order to identify YBX1-dependent targets that are modulated upon changing the levels of endogenous tRFs, we used transient transfection of antisense locked-nucleic acids (LNAs) against tRFAsp, tRFGly, tRFGlu, and tRFTyr followed by microarray profiling. Synthetic antisense locked-nucleic acids (LNAs) targeting the YBX1 binding site on tRFAsp, tRFGly, tRFGlu, and tRFTyr were transfected into control and YBX1-knockdown cells to identify YBX1-dependent targets that are modulated due to tRF loss-of-function.