title: Determining exon connectivity in complex mRNAs by nanopore sequencing.
abstract: Short-read high-throughput RNA sequencing, though powerful, is limited in its ability to directly measure exon connectivity in mRNAs that contain multiple alternative exons located farther apart than the maximum read length. Here, we use the Oxford Nanopore MinION sequencer to identify 7,899 'full-length' isoforms expressed from four Drosophila genes, Dscam1, MRP, Mhc, and Rdl. These results demonstrate that nanopore sequencing can be used to deconvolute individual isoforms and that it has the potential to be a powerful method for comprehensive transcriptome characterization.
authors.affiliation: Department of Genetics and Genome Sciences, Institute for Systems Genomics, University of Connecticut Health Center, Farmington, CT, 06030, USA.|Present Address: The Jackson Laboratory for Genomic Medicine, Farmington, CT, 06030, USA.
authors.forename: Mohan T
authors.initials: MT
authors.lastname: Bolisetty
authors.affiliation: Department of Genetics and Genome Sciences, Institute for Systems Genomics, University of Connecticut Health Center, Farmington, CT, 06030, USA.
authors.forename: Gopinath
authors.initials: G
authors.lastname: Rajadinakaran
authors.affiliation: Department of Genetics and Genome Sciences, Institute for Systems Genomics, University of Connecticut Health Center, Farmington, CT, 06030, USA. graveley@uchc.edu.
authors.forename: Brenton R
authors.initials: BR
authors.lastname: Graveley
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D000818:Animals; D015815:Cell Adhesion Molecules; D004330:Drosophila; D029721:Drosophila Proteins / Q000235:genetics; D005091:Exons / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D009218:Myosins / Q000235:genetics; D058608:Nanopores; D019006:Neural Cell Adhesion Molecules / Q000235:genetics; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D011963:Receptors, GABA-A / Q000235:genetics; D059467:Transcriptome / Q000235:genetics
chemical_list: D018528:ATP-Binding Cassette Transporters; D015815:Cell Adhesion Molecules; D029721:Drosophila Proteins; C491736:Dscam1 protein, Drosophila; C495706:MRP protein, Drosophila; D019006:Neural Cell Adhesion Molecules; D020033:Protein Isoforms; D012333:RNA, Messenger; C082030:Rdl protein, Drosophila; D011963:Receptors, GABA-A; C560638:myosin-18, Drosophila; D009218:Myosins