title: Analysis of remnant reticulocyte mRNA reveals new genes and antisense transcripts expressed in the human erythroid lineage.
abstract: OBJECTIVE
  We studied the gene expression profile of human purified reticulocytes to provide a transcriptional basis for the study of erythroid biology, differentiation and hematologic disorders.
  
  
  METHODS
  We screened highly purified blood reticulocytes from ten healthy adult volunteers. We chose a modified protocol of serial analysis of gene expression (SAGE), the serial analysis of downsized extracts (SADE).
  
  
  RESULTS
  Data analysis revealed that 64% of gene signatures (tags) matched with known genes; mainly hemoglobin. In addition to the abundant globin mRNA, SAGE analysis identified previously described genes and new transcripts. In reticulocytes, which are poor in mRNA, we also identified 9% of EST and 27% of tags that did not match with any known genes. Mining our data, 70% of the unknown tags and 39% of tags identifying EST were found to be specific to the reticulocyte. We demonstrated the presence of a mRNA that matched with the reverse sequence of the hemoglobin b (HBB) transcript.
  
  
  CONCLUSIONS
  This is the first description of an antisense transcript of the human HBB gene suggesting regulation by way of sense-antisense pairing. The well-characterized genes found in the SAGE library were genes specific to the blood cell lineage, housekeeping genes and, interestingly, genes not previously described in the reticulocyte. Furthermore the study provides markers of the erythroid lineage regulated during the differentiation process as observed in in vitro experiments.
authors.affiliation: Institut de Génétique Humaine UPR CNRS 1142, 34396 Montpellier cedex 5, France.
authors.forename: Béatrice
authors.initials: B
authors.lastname: Bonafoux
authors.forename: Mireille
authors.initials: M
authors.lastname: Lejeune
authors.forename: David
authors.initials: D
authors.lastname: Piquemal
authors.forename: Ronan
authors.initials: R
authors.lastname: Quéré
authors.forename: Aurélie
authors.initials: A
authors.lastname: Baudet
authors.forename: Léa
authors.initials: L
authors.lastname: Assaf
authors.forename: Jacques
authors.initials: J
authors.lastname: Marti
authors.forename: Patricia
authors.initials: P
authors.lastname: Aguilar-Martinez
authors.forename: Thérèse
authors.initials: T
authors.lastname: Commes
mesh_terms: D000328:Adult; D001798:Blood Proteins / Q000235:genetics; D002478:Cells, Cultured / Q000378:metabolism; D003199:Computer Systems; D015672:Erythroid Precursor Cells / Q000737:chemistry; D020224:Expressed Sequence Tags; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D005914:Globins / Q000235:genetics; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D016133:Polymerase Chain Reaction; D016372:RNA, Antisense / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification; D012156:Reticulocytes / Q000737:chemistry*
chemical_list: D001798:Blood Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D005914:Globins