title: RNA-seq analysis of single bovine blastocysts.
abstract: BACKGROUND
  Use of RNA-Seq presents unique benefits in terms of gene expression analysis because of its wide dynamic range and ability to identify functional sequence variants. This technology provides the opportunity to assay the developing embryo, but the paucity of biological material available from individual embryos has made this a challenging prospect.
  
  
  RESULTS
  We report here the first application of RNA-Seq for the analysis of individual blastocyst gene expression, SNP detection, and characterization of allele specific expression (ASE). RNA was extracted from single bovine blastocysts (n = 5), amplified, and analyzed using high-throughput sequencing. Approximately 38 million sequencing reads were generated per embryo and 9,489 known bovine genes were found to be expressed, with a high correlation of expression levels between samples (r > 0.97). Transcriptomic data was analyzed to identify SNP in expressed genes, and individual SNP were examined to characterize allele specific expression. Expressed biallelic SNP variants with allelic imbalances were observed in 473 SNP, where one allele represented between 65-95% of a variant's transcripts.
  
  
  CONCLUSIONS
  This study represents the first application of RNA-seq technology in single bovine embryos allowing a representation of the embryonic transcriptome and the analysis of transcript sequence variation to describe specific allele expression.
authors.affiliation: Department of Animal Science, University of California, Davis, Davis, CA, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Chitwood
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Rincon
authors.forename: German G
authors.initials: GG
authors.lastname: Kaiser
authors.forename: Juan F
authors.initials: JF
authors.lastname: Medrano
authors.forename: Pablo J
authors.initials: PJ
authors.lastname: Ross
mesh_terms: D000483:Alleles; D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D002417:Cattle; D005260:Female; D008297:Male; D058977:Molecular Sequence Annotation; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D017423:Sequence Analysis, RNA; D012727:Sex Characteristics; D059467:Transcriptome