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Transcriptional termini of genes on chromosomes 21-22 (RACE mapping)

ABSTRACT: The purpose of these experiments was to determine the 5' and 3' transcriptional termini of genes on chromosomes 21-22. Towards this end, we first carried out 5’ and 3’ RACE reactions from 1193 exons from 492 genes in these chromosomes using RNA isolated from 11 normal tissues and 5 cell lines. An average of 3 to 4 RACE reactions (5’ and 3’) was carried out from each gene. The products of the RACE reactions were analyzed by hybridization to tiling arrays interrogating the non-repeat portions of these chromosomes at 17 nucleotide resolution. A total of 26,688 RACE reactions were performed and pooled to be placed on to 1020 tiling arrays For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf

ORGANISM(S): Homo sapiens

PROVIDER: GSE17600 | GEO | 2009/08/13

REPOSITORIES: GEO

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Efficient targeted transcript discovery via array-based normalization of RACE libraries

Project description:RACE (Rapid Amplification of cDNA Ends) is a widely used approach for transcript identification. However, the dynamic range in the population of RACE transcript isoforms may be very large, and random clone selection -the typical approach- may be ineffective in sampling the different transcript species present in the population. Here, we describe an effective RACE sampling strategy. The products of the RACE reaction are hybridized onto high-density tiling arrays, and the exons detected are then used to delineate a series of RT-PCR reactions, through which the original RACE mixture is segregated into a number of simpler RT-PCR reactions. These are independently cloned, and randomly selected clones are sequenced. This approach is superior to the direct cloning and sequencing of the RACE products: it specifically targets novel transcripts, and often leads to the overall normalization of their abundances. We indeed show theoretically that this strategy leads to a very efficient sampling of the novel transcript species associated to annotated loci. In a pilot experiment, we used this approach to discover many novel transcripts for a few otherwise well-characterized protein coding genes. Finally we investigate how this strategy can be multiplexed for large-scale transcript discovery by high-density pooling of RACE reactions prior to hybridization. Our results indicate that through the interrogation of a limited number of exons per gene on a limited number of cell types, it is possible to recover a large fraction of the transcript diversity associated to protein coding loci. These loci, however, could be occupying a much larger genomic space than previously expected, implying that efficient multiplexing requires non-trivial pooling optimization.

2008-05-25 | GSE11433 | GEO

Transcriptional termini of genes on chromosomes 21-22 (RNA mapping)

Project description:The purpose of these experiments was to determine the 5' and 3' transcriptional termini of genes on chromosomes 21-22. Towards this end, we mapped the poly A + RNA isolated from 12 normal tissues and four cell lines. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf polyA+ RNAs from 11 tissues (Brain Frontal Lobe, Brain Hippocampus, Brain Hypothalamus, Cerebellum, Ovary, Placenta, Prostate, Testis, Fetal Kidney, Fetal Spleen, Fetal Thymus; all from BD Clontech) and 5 cell lines (GM06990, HeLaS3, HepG2, K562, tert-BJ) using the BD SMARTTM RACE cDNA amplification kit (BD Clontech Cat. No.634914). A single array with no replicates was run for each sample with a pool of ~23-24 RACE reactions per sample.

2010-05-15 | E-GEOD-17358 | biostudies-arrayexpress

Obesity and Race Alter Gene Expression in Skin

Project description:We studied differences in epithelial thickness by histology and gene expression by Affymetrix gene arrays and PCR in the skin/fat of 10 obese (BMI 35-50) and 10 normal weight (BMI 18.5-26.9) postmenopausal women paired by age and race

2020-08-26 | GSE151839 | GEO

Transcriptomic analysis of race L Botryococcus braunii AC768 revealed a race L-specific protein resembling the lycopaoctaene synthase

Project description:The green algal Botryococcus braunii (Chlorophyte) is known for accumulating high levels of hydrocarbons that are a useful alternative to fossil fuels. B. braunii is categorized into three groups based on types of their accumulated hydrocarbons: alkadiene/triene in race A, botryococcenes in race B, and lycopadiene in race L. Transcriptomic studies in race A and race B have discovered tremendous information related to the genes encoding proteins involved in hydrocarbon biosynthesis. However, transcriptome of race L has not been reported. In this study, we report a transcriptome of race L B. braunii AC768 through the de novo assembly using Hiseq platform. Our analyses indicate that photosynthesis and protein biosynthesis are the most abundantly transcribed in actively growing race L B. braunii. We show that the transcriptome of race L shares similar amounts (~20%) of mutual best-hits with that of race A or race B. Sequence homologous analyses indicate that enzymes involved in squalene and phytoene biosynthesis are well separated into geranyl-diphosphate synthase, farnesyl-diphosphate synthase, geranylgeranyl-diphosphate synthase, phytoene synthase, and squalene synthase. Both B. braunii specific enzymes botryococcene synthase SSL3 and lycopaoctaene synthase LOS are found to form distinctive subgroups in the group of squalene synthase. One of the ESTs in AC768 transcriptome that falls into the subgroup with LOS and shares >88% identity with that of LOS. Together, our results show that SSL and LOS are unique to race B and race L B. braunii subspecies, respectively. We propose that phytoene synthase in race L shares higher homolog to squalene synthase than phytoene synthase in other algae.

2019-11-21 | GSE96585 | GEO

Extension of 5’ and 3’ ends of human lncRNA transcripts by RACE-coupled long-read high-throughput sequencing (RACE-seq)

Project description:Long non-coding RNAs (lncRNA) constitute a large fraction of mammalian transcriptomes that still remains unexplored, mainly due to the lack of comprehensive, high-quality lncRNA annotation that limits the possibility to fully explore their functional capacity. We have developed RACE-seq, an experimental workflow based on RACE (Rapid Amplification of cDNA Ends) and long read RNA sequencing, aimed at both rare isoform discovery and better definition of gene boundaries. We applied 3â and 5â RACE-seq on 398 low-expressed GENCODE v7 lncRNA genes in seven human tissues (brain, testis, heart, kidney, liver, lung and spleen). The sequences obtained led to the discovery of 2,641 on-target, previously unknown alternative transcripts. Novel isoforms extended 60% of the 398 targeted lncRNA loci further in either 5' or 3', and often reached genome hallmarks typical of gene boundaries. In parallel, we used nested RACE-seq, and confirmed that nested RACE-seq has overwhelmingly better sensitivity than its standard counterpart.

2016-06-01 | E-MTAB-3912 | biostudies-arrayexpress

High-throughput mapping of he promoters of the mouse olfactory receptor genes reveals a new type of mammalian promoters

Project description:We map the transcription start sites of 1085 murine olfactory receptor genes and analyze putative promoters The bar files contain MAT analysis of RLM-RACE products hybridized to a custom olfactory genome tiling array. RNA from the olfactory epithelia of adult mice was prepared by RLM-RACE. Olfactroy receptor transcripts were amplified by degenerate priming and hybridized to tiling arrays to map 5' transcript structure.

2011-05-31 | E-GEOD-26373 | biostudies-arrayexpress

Transcriptome Analysis of Tomato Genotypes in Response to Bacterial spot (Xanthomonas perforans) Race T4 Inoculation

Project description:Purpose: Investigate genes associated to resistance of Xanthomonas perforans race T4 in tomato line with different resistance level Methods: Resistant and susceptible tomato breeding lines were subjected to the inoculation with Xanthomonas perforans race T4 followed by sample collection at 48 hpi and RNA-seq analysis for screening differential expressed genes associated with inoculation of pathogen. Results: Revealed gene expression profiles associated disease resistance and susceptiblilty.

2019-08-02 | GSE135232 | GEO

Role of DNA methylation in FOXA1 cell type-specific recruitment to chromatin

Project description:CTCF, H2AFZ and FOXA1 genomic recruitment sites were determined using ChIP-chip while MeDIP-chip was used to monitor DNA methylation levels. Amplified and labeled DNA was hybridized to Affymetrix tiling arrays covering human chromosomes 8, 11 and 12. Cells used in this study are: MCF7 breast cancer cells, LNCaP prostate cancer cells, MDA-MB-231 breast cancer cells stably transfected with a FOXA1 expression vector (MDA231-FOXA1) or the empty control plasmid (MDA231-CTRL). H3K4me2 genomic distribution was determined using ChIP-chip. Amplified and labeled DNA was hybridized to Affymetrix tiling arrays covering human chromosomes 8, 11 and 12. Cells used in this study are MDA-MB-231 breast cancer cells stably transfected with a FOXA1 expression vector (MDA231-FOXA1) or the empty control plasmid (MDA231-CTRL).

2011-01-06 | GSE21513 | GEO

Resistant response of leaves of upland cotton line Im216 to Xanthomonas campestris pv. malvacearum race 1

Project description:Gene expression profiling of upland cotton line Im216 to inoculation with Xanthomonas campestris pv. malvacearum race 1. Fifth or sixth leaves of the bacterial blight-resistant cotton line Im216, which had been grown in a plant growth chamber, were infiltrated with a suspension of about 5x10^6 colony-forming units ml^-1 of Xanthomonas campestris pv. malvacearum race 1 in sterile saturated CaCO3 solution or were not inoculated (control). Keywords: Time-course

2006-01-01 | GSE2851 | GEO

Phytophthora sojae, race 2, infection of soybean hypocotyls

Project description:Examination of soybean hypocotyls, G. max cv. Harosoy (Rps7), at 3, 6, 12, 24 and 48 hours after inoculation with P. sojae, race 2, isolate P6497 Patterns of Gene Expression Upon Infection of Soybean Plants by Phytophthora sojae. P. Moy, D. Qutob, B. P. Chapman, I. Atkinson, and M. Gijzen. Pages 1051-1062. Publication no. M-2004-0728-01R. Molecular Plant-Microbe Interactions, October 2004, Volume 17, Number 10. Keywords: time-course

2004-09-14 | GSE1352 | GEO

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