Comprehensive Serial Analysis of Gene Expression of the Cervical Epithelium Transcriptome
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE6252 | GEO | 2008/02/19
SECONDARY ACCESSION(S): PRJNA99681
REPOSITORIES: GEO
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Project description:In this study, we sequenced 691,390 SAGE tags from four libraries. Cervical L-SAGE libraries N1, N2, C1, and C2 were sequenced to 165,624, 181,224, 173,534, and 171,008 tags, respectively. Duplicate ditags were eliminated from analysis resulting in 136,276, 139,656, 154,828 and 136,386 useful tags respectively and a total of 24 058 unique tags. 15,438 of the unique tags mapped to annotated UniGene identifiers. We characterized the transcriptome of normal cervical tissue and evaluated the highly expressed genes in terms of tissue specificity, conserved expression among the normal libraries and their altered expression in CIN III lesions. Keywords: Cervical Epithelium, Long SAGE Four Long SAGE libraries were created from cervical epithelium biopsies. Two were CIN III and two were normal cervical tissue.
2008-02-18 | E-GEOD-6252 | biostudies-arrayexpress
Project description:The analysis of differentially expressed genes is a powerful approach to elucidate the genetic mechanisms underlying the morphological and evolutionary diversity among serially homologous structures, both within the same organism (e.g., hand vs. foot) and between different species (e.g., hand vs. wing). In the developing embryo, limb-specific expression of Pitx1, Tbx4, and Tbx5 regulates the determination of limb identity. However, numerous lines of evidence, including the fact that these three genes encode transcription factors, indicate that additional genes are involved in the Pitx1-Tbx hierarchy. To examine the molecular distinctions coded for by these factors, and to identify novel genes involved in the determination of limb identity, we have used Serial Analysis of Gene Expression (SAGE) to generate comprehensive gene expression profiles from intact, developing mouse forelimbs and hindlimbs. To minimize the extraction of erroneous SAGE tags from low-quality sequence data, we used a new algorithm to extract tags from -analyzed sequence data and obtained 68,406 and 68,450 SAGE tags from forelimb and hindlimb SAGE libraries, respectively. We also developed an improved method for determining the identity of SAGE tags that increases the specificity of and provides additional information about the confidence of the tag-UniGene cluster match. The most differentially expressed gene between our SAGE libraries was Pitx1. The differential expression of Tbx4, Tbx5, and several limb-specific Hox genes was also detected; however, their abundances in the SAGE libraries were low. Because numerous other tags were differentially expressed at this low level, we performed a 'virtual' subtraction with 362,344 tags from six additional nonlimb SAGE libraries to further refine this set of candidate genes. This subtraction reduced the number of candidate genes by 74%, yet preserved the previously identified regulators of limb identity. This study presents the gene expression complexity of the developing limb and identifies candidate genes involved in the regulation of limb identity. We propose that our computational tools and the overall strategy used here are broadly applicable to other SAGE-based studies in a variety of organisms. Keywords: other
2003-07-16 | GSE504 | GEO
Project description:This study characterizes the expressed transcripts of 15 intact tissues in mice by using the serial analysis of gene expression (SAGE) strategy which indicates the relative level of expression for each transcript matched to the tag. Keywords: double-strand cDNA , serial analysis of gene expression (SAGE) , ditags
2007-04-03 | GSE5915 | GEO
Project description:Müller cells are the principal glial cells in the retina. Alterations in Müller cell behaviour are observed in retinal tissue from patients with proliferative diabetic retinopathy. The purpose of this study was to compare gene and protein expression profile of normal human Müller cells (NHMC) with two spontaneously human Müller cell lines generated from type 1 (HMCL-I) and type 2 (HMCL-II) diabetic donors using Serial Analysis Gene Expression (SAGE). Approximatively 50 000 tags were sequenced for each of the three SAGE libraries. Identification of the transcripts was obtained by matching the 15bp (CATG + 11bp) with the UniGene and GenBank databases. Classification of the genes was based upon the updated information of the genome directory found at the NCBI website. SAGE allowed us to characterize the entire transcriptome of human Müller cells and to compare these data with those from Müller cells of type 1 and 2 diabetic donors. Keywords: Müller cells, SAGE, HMCL-I, HMCL-II, expression profile comparison
2006-08-12 | GSE3118 | GEO
Project description:Transcriptomes fiber and ovules were compared by applying serial analysis of gene expression (SAGE). Keywords: Tissue Comparison We constructed three SAGE libraries and sequenced 57321, 64188, and 69104 tags from fiber, Xu-142 ovule (ovule) and fl mutant ovules (fl) respectively of Upland Cotton, Gossypium hirsutum L. cv. Xu-142.
2007-02-17 | E-GEOD-6103 | biostudies-arrayexpress
Project description:The analysis of differentially expressed genes is a powerful approach to elucidate the genetic mechanisms underlying the morphological and evolutionary diversity among serially homologous structures, both within the same organism (e.g., hand vs. foot) and between different species (e.g., hand vs. wing). In the developing embryo, limb-specific expression of Pitx1, Tbx4, and Tbx5 regulates the determination of limb identity. However, numerous lines of evidence, including the fact that these three genes encode transcription factors, indicate that additional genes are involved in the Pitx1-Tbx hierarchy. To examine the molecular distinctions coded for by these factors, and to identify novel genes involved in the determination of limb identity, we have used Serial Analysis of Gene Expression (SAGE) to generate comprehensive gene expression profiles from intact, developing mouse forelimbs and hindlimbs. To minimize the extraction of erroneous SAGE tags from low-quality sequence data, we used a new algorithm to extract tags from -analyzed sequence data and obtained 68,406 and 68,450 SAGE tags from forelimb and hindlimb SAGE libraries, respectively. We also developed an improved method for determining the identity of SAGE tags that increases the specificity of and provides additional information about the confidence of the tag-UniGene cluster match. The most differentially expressed gene between our SAGE libraries was Pitx1. The differential expression of Tbx4, Tbx5, and several limb-specific Hox genes was also detected; however, their abundances in the SAGE libraries were low. Because numerous other tags were differentially expressed at this low level, we performed a 'virtual' subtraction with 362,344 tags from six additional nonlimb SAGE libraries to further refine this set of candidate genes. This subtraction reduced the number of candidate genes by 74%, yet preserved the previously identified regulators of limb identity. This study presents the gene expression complexity of the developing limb and identifies candidate genes involved in the regulation of limb identity. We propose that our computational tools and the overall strategy used here are broadly applicable to other SAGE-based studies in a variety of organisms. Keywords: other
2003-07-16 | E-GEOD-504 | biostudies-arrayexpress
Project description:Serial analysis of gene expression (SAGE) was used to identify and quantify all expressed cerebellar genes in the adult (P92; GSM17430) and aged (P810; GSM17226) C57BL/6J mouse cerebellum. A "closest-neighbor" algorithm was used to differentiate low abundance tags from possible sequencing errors in both libraries. Unique tags were categorized into four groups: (1) novel genes; (2) ESTs; (3) RIKEN, KIA, and hypothetical genes; and (4) known genes. Known genes were further subdivided into functional categories based on the gene ontology classification, using a web-based program developed in this laboratory (MmSAGEClass). Comparison of adult and aged cerebellar libraries revealed several genes that were differentially expressed, including growth hormone and prolactin, both of which were markedly decreased in the aged cerebellum. In addition, several tags showing differential expression were not identified in the Unigene database and are likely to represent novel genes. The present SAGE data on the aged cerebellar transcriptome may reveal candidate genes involved in the aging process. Keywords: other
2004-10-26 | GSE1090 | GEO
Project description:Cardiac fibroblasts were isolated from 3-month old C57BL/6 mice. RNA was prepared from quiescent, serum-starved cardiac fibroblasts in passage 15. A total number of 118654 tags was sequenced. After removal of duplicate ditags and linker-derived sequences 110169 tags remained.
2010-12-01 | GSE25757 | GEO
Project description:Biomphalaria glabrata infection by the Schistosoma mansoni free-swimming miracidium and its subsequent development to the parasitic sporocyst stage is critical to establishment of viable infections and triggers a variety of physiological, biochemical and molecular changes. Here, we describe a genome-wide analysis of the S. mansoni miracidium and developing sporocyst. Keywords: life-cycle, development, host-interaction We generated transcriptomic profiles of the developing larval stages of Schistosoma mansoni using long serial analysis of gene expression (LongSAGE). Five cDNA libraries were constructed from miracidia and in vitro cultured 6- and 20-day old sporocysts maintained in sporocyst medium (SM) or in SM conditioned by previous cultivation with cells of the B. glabrata embryonic (Bge) cell line. From five libraries, 314,799 SAGE tags were sequenced and resulted in a total of 21,440 unique sequence tags. A total of 254 tags were differentially expressed during “conditioned” development and 236 tags were differentially expressed during “un-conditioned” development. In addition, 53 tags were found to be differentially expressed between 6-day conditioned and unconditioned sporocysts and 42 tags between 20-day conditioned and unconditioned sporocysts.
2009-03-09 | E-GEOD-9722 | biostudies-arrayexpress
Project description:This study characterizes the expressed transcripts of 15 intact tissues in mice by using the serial analysis of gene expression (SAGE) strategy which indicates the relative level of expression for each transcript matched to the tag. Keywords: double-strand cDNA , serial analysis of gene expression (SAGE) , ditags Total RNA was isolated from tissues by using the RNA extraction kit (TRIzol Reagent, Invitrogen Canada Inc., Burlington, ON). Approximately 5 µg of mRNA was extracted with Oligotex mRNA Mini Kit (Qiagen Inc., Mississauga, ON). The SAGE method was performed as previously described (Velculescu et al., 1995; St-Amand et al., 2001). In brief, double-strand cDNA was synthesized from the mRNA using a biotinylated (T)18 primer and cDNA synthesis kit (Invitrogen Canada Inc.). The cDNA libraries were digested with the restriction enzyme NlaIII (New England Biolabs Inc., Pickering, ON). The 3'-terminal cDNA fragments were captured using streptavidin-coated magnetic beads (Dynal, Biotech LLC, Brown Deer, WI). After ligation of 2 annealed linker pairs, the cDNA fragments were digested with BsmFI (New England Biolabs Inc.). The blunting kit from Takara Bio Inc. (Otsu, Japan) was used for the blunting and ligation of the two tag populations. The resulting ligation products were amplified by PCR and digested with NlaIII. The band containing the ditags was extracted from the 12% polyacrylamid gel. Using T4 ligase (Invitrogen Canada Inc.), the ditags were self-ligated to form concatemers that were cloned into SphI site of pUC19. White colonies were screened by PCR and agarose gel to select long inserts for automated sequencing (Applied Biosystems 3730, Foster City, CA). The sequence and occurrence of each tag were analyzed by the SAGEana program, which is a new version of SAGEparser.pl (Dinel et al., 2005).
2007-04-03 | E-GEOD-5915 | biostudies-arrayexpress