title: Polony multiplex analysis of gene expression (PMAGE) in mouse hypertrophic cardiomyopathy.
abstract: We describe a sensitive mRNA profiling technology, PMAGE (for "polony multiplex analysis of gene expression"), which detects messenger RNAs (mRNAs) as rare as one transcript per three cells. PMAGE incorporates an improved ligation-based method to sequence 14-nucleotide tags derived from individual mRNA molecules. One sequence tag from each mRNA molecule is amplified onto a separate 1-micrometer bead, denoted as a polymerase colony or polony, and about 5 million polonies are arrayed in a flow cell for parallel sequencing. Using PMAGE, we identified early transcriptional changes that preceded pathological manifestations of hypertrophic cardiomyopathy in mice carrying a disease-causing mutation. PMAGE provided a comprehensive profile of cardiac mRNAs, including low-abundance mRNAs encoding signaling molecules and transcription factors that are likely to participate in disease pathogenesis.
authors.affiliation: Cardiovascular Division, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Jae Bum
authors.initials: JB
authors.lastname: Kim
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Porreca
authors.forename: Lei
authors.initials: L
authors.lastname: Song
authors.forename: Steven C
authors.initials: SC
authors.lastname: Greenway
authors.forename: Joshua M
authors.initials: JM
authors.lastname: Gorham
authors.forename: George M
authors.initials: GM
authors.lastname: Church
authors.forename: Christine E
authors.initials: CE
authors.lastname: Seidman
authors.forename: J G
authors.initials: JG
authors.lastname: Seidman
mesh_terms: D000818:Animals; D002312:Cardiomyopathy, Hypertrophic / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology; D018076:DNA, Complementary; D005355:Fibrosis / Q000235:genetics / Q000473:pathology; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D015723:Gene Library; D006352:Heart Ventricles / Q000378:metabolism; D051379:Mice; D009154:Mutation; D009200:Myocardial Contraction; D009206:Myocardium / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013698:Templates, Genetic; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D024723:Ventricular Myosins / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D014157:Transcription Factors; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains