Project description:A vast portion of the mammalian genome is transcribed as long non-coding RNAs (lncRNAs) acting in the cytoplasm with largely unknown functions. Surprisingly, lncRNAs have been shown to interact with ribosomes, encode uncharacterized proteins, or act as ribosome sponges. These functions still remain mostly undetected and understudied owing to the lack of efficient tools for genome-wide simultaneous identification of ribosome-associated lncRNAs and peptide-producing lncRNAs. Here we present AHARIBO, a method for the detection of lncRNAs either untranslated, but associated with ribosomes, or encoding small peptides. Using AHARIBO in mouse embryonic stem cells during neuronal differentiation, we isolated ribosome-protected RNA fragments, translated RNAs and corresponding de novo synthesized polypeptides. Besides identifying mRNAs under active translation and associated ribosomes, we found and distinguished lncRNAs acting as ribosome sponges or encoding micropeptides, laying the ground for a better functional understanding of hundreds lncRNAs.
Project description:To understand the role of long non-coding RNAs and interaction with coding RNAs in bladder urothelial cell carcinoma (BUCC), we performed genome-wide screening long non-coding RNAs and coding RNAs expression on primary BUCC tissues and normal tissues using long non-coding RNA array (Agilent plateform (GPL13825). By comparing these two groups, significantly differentially expressed lncRNAs and coding RNAs were identified. We further identifed a subset of long noncoding RNAs and their correlation with neighboring coding genes using bioinformatic tools. This analysis provides foundamental understaning of transcriptomic landscape changing during bladder carcinogenesis.
Project description:Total RNA was isolated from WBCs. For the analysis of genome-wide expression differences in small non-coding RNAs (sncRNAs) and long-coding and non-coding RNAs (mRNAs and lncRNAs), NGT and GDM pregnant women were selected. Twenty-nine GDM-associated mature micro-RNAs (miRNAs) with increased expression and one hundred sixty-three mRNAs with reduced expression associated with GDM were found (P<0.05 and FDR<0.1).
Project description:To understand the role of long non-coding RNAs and interaction with coding RNAs in bladder urothelial cell carcinoma (BUCC), we performed genome-wide screening long non-coding RNAs and coding RNAs expression on primary BUCC tissues and normal tissues using long non-coding RNA array (Agilent plateform (GPL13825). By comparing these two groups, significantly differentially expressed lncRNAs and coding RNAs were identified. We further identifed a subset of long noncoding RNAs and their correlation with neighboring coding genes using bioinformatic tools. This analysis provides foundamental understaning of transcriptomic landscape changing during bladder carcinogenesis. 12 BUCC primary tumors and 3 normal tissues were used for long noncoding RNA array experiments which including long non-coding RNAs and coding RNAs. The differential expression of subset of long noncoding RNAs and their interaction with coding RNAs in BUCC compared with normal tissue will be identified with comtational analysis.
Project description:It has been a long debate whether the 98% “non-coding” fraction of human genome can encode functional proteins besides a “random noise” of translation. We used our established translatome sequencing (RNC-seq) to analyze human cells and found that up to 3330 long non-coding RNAs (lncRNAs) were bound to ribosomes and thus might be translated into proteins (with more than 50 amino acids). These new protein-coding genes distributed universally in all human chromosomes. We then used various experimental methods including mass spectrometry, immunoblotting, subcellular localization and phenotype assessments to verify the existence of such a hidden human proteome encoded by purported lncRNAs that can express functional proteins. These new proteins deviate from the canonical proteins in various physical and chemical properties, and emerged mostly in primates during evolution. In sum, we experimentally evidenced a hidden human functional proteome encoded by purported lncRNAs, suggesting that the human genome has to be systematically re-annotated.
Project description:It has been a long debate whether the 98% “non-coding” fraction of human genome can encode functional proteins besides a “random noise” of translation. We used our established translatome sequencing (RNC-seq) to analyze human cells and found that up to 3330 long non-coding RNAs (lncRNAs) were bound to ribosomes and thus might be translated into proteins (with more than 50 amino acids). These new protein-coding genes distributed universally in all human chromosomes. We then used various experimental methods including mass spectrometry, immunoblotting, subcellular localization and phenotype assessments to verify the existence of such a hidden human proteome encoded by purported lncRNAs that can express functional proteins. These new proteins deviate from the canonical proteins in various physical and chemical properties, and emerged mostly in primates during evolution. In sum, we experimentally evidenced a hidden human functional proteome encoded by purported lncRNAs, suggesting that the human genome has to be systematically re-annotated.
Project description:In order to understand the role of long noncoding RNAs (lncRNAs) and their interaction with coding RNAs in esophageal sqaumous cell cancer (ESCC), we performed genome-wide screening of the expression of lncRNAs and coding RNAs from primary ESCC tissue and adjacent normal tissue using Agilent SurePrint G3 Human GE 8x60K Microarray. By comparing ESCC tissues and matched normal tissues, differentially expressed lncRNAs and coding RNAs were identified and confirmed with PCR and other independent studies. We further identified a subset of co-located and co-expressed lncRNAs and coding RNAs using bioinformatic tools and the analysis suggested that a subset of lncRNAs may influence nearby genes involved in the genesis of ESCC. Four pairs of ESCC primary tumors and adjacent normal tissues were used for genome-scale microarray experiments, which included long noncoding RNAs and coding RNAs. Selected lncRNAs expressed in the experiment were validated on independent matched-pair samples with PCR method.
Project description:Long non-coding RNAs (lncRNAs) comprise a diverse class of transcripts that structurally resemble mRNAs but do not encode proteins. Recent genome-wide studies in human and mouse have annotated lncRNAs expressed in cell lines and adult tissues, but a systematic analysis of lncRNAs expressed during vertebrate embryogenesis has been elusive. To identify lncRNAs with potential functions in vertebrate embryogenesis, we performed a time series of RNA-Seq experiments at eight stages during early zebrafish development. We reconstructed 56,535 high-confidence transcripts in 28,912 loci, recovering the vast majority of expressed RefSeq transcripts, while identifying thousands of novel isoforms and expressed loci. We defined a stringent set of 1,133 non-coding multi-exonic transcripts expressed during embryogenesis. These include long intergenic ncRNAs (lincRNAs), intronic overlapping lncRNAs, exonic antisense overlapping lncRNAs, and precursors for small RNAs (sRNAs). Zebrafish lncRNAs share many of the characteristics of their mammalian counterparts: relatively short length, low exon number, low expression, and conservation levels comparable to introns. Subsets of lncRNAs carry chromatin signatures characteristic of genes with developmental functions. The temporal expression profile of lncRNAs revealed two novel properties: lncRNAs are expressed in narrower time windows than protein-coding genes and are specifically enriched in early-stage embryos. In addition, several lncRNAs show tissue-specific expression and distinct subcellular localization patterns. Integrative computational analyses associated individual lncRNAs with specific pathways and functions, ranging from cell cycle regulation to morphogenesis. Our study provides the first comprehensive identification of lncRNAs in a vertebrate embryo and forms the foundation for future genetic, genomic and evolutionary studies. ChIP-Seq for H3K4me3 and H3K27me3 at zebrafish shield stage.
Project description:Small RNAs target their complementary chromatin regions for gene silencing through nascent long non-coding RNAs (lncRNAs). In programmed DNA elimination of the ciliated protozoan Tetrahymena, the interaction between Piwi-associated small RNA (scnRNAs) and the lncRNA transcripts from the somatic genome has been proposed to induce target-directed scnRNA degradation (TDSD) and scnRNAs not targeted for TDSD later target the germline-limited sequences for DNA elimination. In this study, we show that the SUMO E3 ligase Ema2 is required for the accumulation of lncRNAs from somatic genome, and thus for TDSD and completing DNA elimination to make viable sexual progeny. Ema2 interacts with the SUMO E2 conjugating enzyme Ubc9 and enhances SUMOylation of the transcription regulator Spt6. We further show that Ema2 promotes the association of Spt6 and RNA polymerase II to chromatin. These results suggest that Ema2-directed SUMOylation actively promotes the lncRNA transcription that is a prerequisite for communication between the genome and small RNAs.
Project description:Total RNA sequencing was performed on whole blood samples from 74 Lupus nephritis (LN) patients and 20 healthy controls. Differential expression analysis and weighted gene co-expression network analysis were performed to characterise expression changes of long non-coding RNAs (lncRNAs) in LN and identify lncRNAs with a key role in disease activity that could be used as potential blood-based biomarkers.