Polony multiplex analysis of gene expression (PMAGE) in mouse hypertrophic cardiomyopathy.
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.
Kim
Jae Bum
JB
Cardiovascular Division, Brigham and Women's Hospital, Boston, MA 02115, USA.
Porreca
Gregory J
GJ
Song
Lei
L
Greenway
Steven C
SC
Gorham
Joshua M
JM
Church
George M
GM
Seidman
Christine E
CE
Seidman
J G
JG
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
D018076:DNA, Complementary; D012333:RNA, Messenger; D014157:Transcription Factors; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains