The expression profiles of 82 novel and 245 known human imsRNAs
ABSTRACT: The expression profiling of 245 known and 82 novel imsRNAs (intermediate-size noncoding RNAs, 50-500nt, which were identified from human fetal brain tissue) across different human fetal tissues (brain, heart, liver, lung and spleen at the 24-week gestation stage), different fetal brain gestational stages (13w, 14w, 20w, and 24w) and two human brain tumor cell lines (SHSY-5Y and U251). The analysis was based on the loop design method, in which all combinations of two samples were simultaneously hybridized to a microarray, each of the two sample labeled with a different fluorescent probe (Cy3 or Cy5).
Project description:The expression profiling of 245 known and 82 novel imsRNAs (intermediate-size noncoding RNAs, 50-500nt, which were identified from human fetal brain tissue) across different human fetal tissues (brain, heart, liver, lung and spleen at the 24-week gestation stage), different fetal brain gestational stages (13w, 14w, 20w, and 24w) and two human brain tumor cell lines (SHSY-5Y and U251). Overall design: The analysis was based on the loop design method, in which all combinations of two samples were simultaneously hybridized to a microarray, each of the two sample labeled with a different fluorescent probe (Cy3 or Cy5).
Project description:We detected 32230 transfrags by Tiling array Analysis Software, of which 5866 were transfrags with unknown function (TUF). These TUFs exhibited distinct chromosome location, expression and conservation patterns. Six developmental and two conditional stages of C. elegans 70~500nt transcriptome. L1~L4, mature adult (MA), male (ML), dauer (DU), heat shocked (HS). GSM591423_CB2007199-L4-1.CEL is Ce25b_MF, GPL5634 is Ce25b_MR, CEL file is okay using GPL5634's cdf.
Project description:Recent advances in genome-wide techniques allowed the identification of thousands of non-coding RNAs with various sizes in eukaryotes, some of which have further been shown to serve important functions in many biological processes. However, in model plant Arabidopsis, novel intermediate-sized ncRNAs (im-ncRNAs) (50~300nt) have very limited information. By using a modified isolation strategy combined with deep-sequencing technology, we identified 838 im-ncRNAs in Arabidopsis globally. More than half (58%) are new ncRNA species, mostly evolutionary divergent. Interestingly, annotated protein-coding genes with 5’-UTR derived novel im-ncRNAs tend to be highly expressed. For intergenic im-ncRNAs, their average abundances were comparable to mRNAs in seedlings, but subsets exhibited significantly lower expression in senescing leaves. Further, intergenic im-ncRNAs were regulated by similar genetic and epigenetic mechanisms as those of protein-coding genes, and some showed developmentally-regulated expression patterns. Large-scale reverse genetic screening showed that the down-regulation of a number of im-ncRNAs resulted in either obvious molecular changes or abnormal developmental phenotypes in vivo, indicating the functional importance of im-ncRNAs in plant growth and development. Together, our results demonstrate that novel Arabidopsis im-ncRNAs are developmentally-regulated and functional components discovered in the transcriptome. Genome-wide maps of Intermediate-size Non-coding RNAs in Arabidopsis
Project description:Deep sequencing of Transcriptional Start Sites (TSS) using 5' CAGE from 82 wild strains from the Drosophila Genetic reference Panel (DGRP) during embryogenesis. Three different developmental stages were assayed: 2-4hrs (stages 5-8), 6-8hrs (stages 10-11) and 10-12hrs (stages 13-14) after laying. 20 line/stage combinations were performed in biological replicates (independent embryo collections), while 5 samples were prepared as technical replicates (independent library preparation from the same RNA).
Project description:Corneal endothelium is composed of a monolayer of corneal endothelial cells (CECs) in the inner layer of cornea, which is essential for maintaining corneal transparency. In order to better characterize CECs in different developmental stages, we profiled mRNA transcriptomes in human fetal and adult corneal endothelium with the goal to identify novel molecular markers in these cells. By comparing CECs with 12 other types of tissues, we identified 245 and 284 signature genes that are highly expressed in fetal and adult CECs, respectively. Functionally, these genes are characteristic of CECs, involving in cell adhesion, proteoglycan and sulfur metabolic process. Importantly, several of these genes are disease target genes in hereditary corneal dystrophies, consistent with their functional significance in CEC physiology. By comparing fetal and adult CECs, we also identified stage-specific markers associated with CEC maturation, such as the activation of the Wnt pathway genes in fetal, but not in adult CECs. Lastly, by immunohistochemistry of ocular tissues, we further confirmed the unique protein expression patterns for Wnt5a, S100A4, S100A6, and IER3, the four novel markers for either fetal or adult CECs. The identification of a new panel of molecular markers for fetal and mature CECs would be very useful for characterizing and quality controlling CECs through ex vivo expansion or stem cell differentiation for cell replacement therapy. mRNA profile between adult and fetal CECs by high-throughput sequencing
Project description:Brain-derived neurotrophic factor (BDNF) is a neurotrophin family member that is highly expressed and widely distributed in the brain. BDNF is critical for neural survival and plasticity both during development and in adulthood, and dysfunction in its signaling may contribute to a number of neurodegenerative disorders. Deep understanding of the BDNF-activated molecular cascade may thus help to find new biomarkers and therapeutic targets. One interesting direction is related to the early phase of BDNF-dependent gene expression regulation, which is responsible for the activation of selective gene programs that lead to stable functional and structural remodeling of neurons. Immediate-early coding genes activated by BDNF are under investigation, but the involvement of the non-coding RNAs is largely unexplored, especially the long non-coding RNAs (lncRNAs). lncRNAs are emerging as key regulators that can orchestrate different aspects of nervous system development, homeostasis, and plasticity, making them attractive candidate markers and therapeutic targets for brain diseases. We used microarray technology to identify differentially expressed lncRNAs in the immediate response phase of BDNF stimulation in a neuronal cell model. Our observations on the putative functional role of lncRNA provide clues to their involvement as master regulators of gene expression cascade triggered by BDNF. Overall design: We used the popular SHSY-5Y cell line as a neuronal cell model for BDNF stimulation. The SHSY-5Y line comprises at least two morphologically and biochemically distinct phenotypes: neuroblastic (N-type) and a low proportion of epithelial-like (S-type) phenotypes. For BDNF treatment on a more homogeneous neuronal cell population, we performed an enrichment procedure based on the different substrate adherence between the two cell phenotypes. The obtained N-enriched population of SHSY-5Y was differentiated by decreasing FBS concentration from 15% to 1.5% and adding 10 μM of retinoic acid for 6 days (the medium was refreshed every 2 days). After 6 days of differentiation, the medium containing 1.5% FBS and RA was removed and substituted with a medium without FBS for two groups of cells. One of these groups was treated with 10 ng/mL of BDNF for 1 h, whereas the second group was not treated and used as a control for the gene expression analysis. Microarray experiments were performed on biological triplicate samples.
Project description:We provided a full spectrum analysis for E. histolytica AGO2-2 associated 27nt small RNAs. Additionally, comparative analysis of small RNA populations from virulent and non-virulent amebic strains indicates that small RNA populations may regulate virulence genes. AGO2-2 bound small RNAs from E. histolytica strain HM-1:IMSS were immunoprecipitated and sequenced using 454 technology. Three independant sequencing runs were perfomed using the same RNA sample. In addition, size selected small RNAs from E. histolytica strain Rahman were sequenced with the same technology. One sequencing run was performed on this sample.
Project description:Study on sequencing biases introduced by library preparation step, namely by ligases. Comparison between standard Illumina protocol and improved High Definition (HD) protocol. Four replicates for N21 (21 random nucleotides), one replicate for N9 (9 random nucleotides), using either standard Illumina protocol or HD protocol
Project description:Chromatin Immunoprecipitation of Centromeric Proteins, CenpA, CenpC or CenpH from cell line BBB, which contains a neocentromere in band 13q33.1<br><br> Processed data files with additional statistics are available for download from <a href="ftp://ftp.ebi.ac.uk/pub/databases/microarray/data/experiment/TABM/E-TABM-245/">ftp://ftp.ebi.ac.uk/pub/databases/microarray/data/experiment/TABM/E-TABM-245/</a>
Project description:Embryo DNA fingerprinting represents an important tool for tracking embryo-specific outcomes after multiple embryo transfer during IVF. The situation in which 2 embryos are transferred and only one implants represents a unique opportunity for the most well-controlled comparison of competent and incompetent embryos. Specifically, this design eliminates all patient-related variables from the comparison of embryos with or without reproductive potential. However, in order to determine which embryo implanted, the investigator must wait until newborn DNA is available upon delivery. This study validates a non-invasive fetal DNA fingerprinting method that reduces the time to identify which embryo implanted by approximately 31 weeks. Thirty-four patients were studied to determine if fingerprinting of fetal DNA extracted from maternal plasma at 9 gestational weeks concurred with the buccal DNA results obtained from the newborn after delivery. This validation required single nucleotide polymorphism (SNP) profiles on each couples’ preimplantation embryos, enriched fetal DNA from maternal plasma at 9 weeks gestation, and newborn DNA obtained from buccal swabs after delivery. The predictions from fetal DNA-based embryo tracking and gender assignments made at 9 weeks gestation were 100% consistent with standardized methods of assessment performed after term delivery. This study demonstrates the first validated fetal DNA fingerprinting method which predicts both gender and which embryo implanted at 9 weeks gestation following multiple embryo transfer. Affymetrix SNP arrays were processed and successfully completed according to the manufacturer's directions on DNA extracted from 136 embryos, 33 parental blood samples, 17 enriched fetal DNA samples and 21 buccal DNA samples.