Single molecule profiling of alternative pre-mRNA splicing.
Alternative pre-messenger RNA splicing is an important mechanism for generating protein diversity and may explain in part how mammalian complexity arises from a surprisingly small complement of genes. Here, we describe "digital polony exon profiling,"a single molecule-based technology for studying complex alternative pre-messenger RNA splicing. This technology allows researchers to monitor the combinatorial diversity of exon inclusion in individual transcripts. A minisequencing strategy provides single nucleotide resolution, and the digital nature of the technology allows quantitation of individual splicing variants. Digital polony exon profiling can be used to investigate the physiological and pathological roles of alternately spliced messenger RNAs, as well as the mechanisms by which these messenger RNAs are produced.
Zhu
Jun
J
Department of Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.
Shendure
Jay
J
Mitra
Robi D
RD
Church
George M
GM
D020106:Acrylamide; D017398:Alternative Splicing; D000818:Animals; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D002461:Cell Line, Transformed; D017362:Cyclic AMP Response Element-Binding Protein; D005091:Exons; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D020033:Protein Isoforms; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins; D055532:SMN Complex Proteins
D017362:Cyclic AMP Response Element-Binding Protein; D018960:Hyaluronan Receptors; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D055532:SMN Complex Proteins; D020106:Acrylamide