MAR 15, 2016 10:00 AM PDT

Gene Expression Analysis Using 3'-RNA Sequencing

Sponsored by: Lexogen, Lexogen
Speaker
  • Assistant Professor, University of Delaware
    Biography
      Behnam Abasht joined University of Delaware's faculty as an Assistant Professor in 2011. He received his PhD in Quantitative and Molecular Genetics in France from INRA-Agrocampus Rennes in 2006. He then completed a postdoctoral fellowship in Quantitative Genetics at Iowa State University. Dr. Abasht was a Research Geneticist and Genomics Project Leader at Perdue Farms, Inc. from 2008 until his appointment at the University of Delaware. His area of research is integrative avian biology with emphasis on fine-mapping and functional characterization of quantitative trait loci (QTL) in chickens.

    Abstract
    10:00AM PT, 12:00PM CT, 1:00PM ET

    RNA sequencing (RNA-seq) has revolutionized the study of gene expression in animals, plants and microorganisms. However, because of its high cost, this technology has been mainly used in experiments with limited number of samples. To examine a cost-effective alternative, we used a method, which confines sequencing to the 3’-end of mRNA and produces just one fragment per transcript, resulting in a dramatic decrease in sequencing cost. Total RNA isolated from chicken adipose tissue samples was used for cDNA library preparation using QuantSeq 3’mRNA-seq library Prep Kit. Sixty-one uniquely indexed cDNA libraries were pooled and sequenced on one lane on the Illumina Hiseq 2500. On average, 2.24 million reads per sample were generated, 90.1% of which were mapped to the chicken reference genome (Ensembl Galgal4). For more than 70% of the genes with detectable expression, we redefined the 3’-end and identified alternative polyadenylation sites within the 3’-untranslated regions. To compare gene expression measures between 3’-RNA-seq and RNA-seq technologies, we used data from a subset of 20 samples that were previously used in a RNA-seq study of feed efficiency. The correlation of the log10(fold-change) for gene expression (high- vs. low-feed efficiency birds) between these two methods was 0.90. In conclusion, 3’-RNA-seq is a cost effective method amenable to global gene expression studies at population-level, e.g., expression QTL (eQTL) mapping.  Also, it allows for accurate detection of the 3’-end of transcripts, enabling verification of the current gene model annotations and global characterization of alternative polyadenylation.

    Learning objectives:

    1) Gain knowledge about 3’-RNA-seqencing and its application for global gene expression studies at population-level
     
    2) Explore how 3’-RNA-seqencing can help improve current gene model annotations and characterize alternative polyadenylation

     

    Show Resources
    You May Also Like
    JAN 23, 2020 9:00 AM PST
    C.E. CREDITS
    JAN 23, 2020 9:00 AM PST
    DATE: January 23, 2020 TIME: 9:00am PST, 12:00pm EST...
    MAY 08, 2020 10:00 AM PDT
    C.E. CREDITS
    MAY 08, 2020 10:00 AM PDT
    DATE: May 8, 2020 TIME: 10:00am PT, 11:00am MT, 1:00pm ET The application of next generation sequencing to interrogate immune repertoires and methods in which these highly complex dataset...
    APR 07, 2020 8:00 AM PDT
    C.E. CREDITS
    APR 07, 2020 8:00 AM PDT
    DATE: April 7, 2020 TIME: 8:00am PT, 11:00am ET This webinar sets out to establish why quality control is key to robust, reliable, reproducible science. We will look at best practice criteri...
    FEB 26, 2020 9:00 AM PST
    C.E. CREDITS
    FEB 26, 2020 9:00 AM PST
    DATE: February 26, 2020 TIME: 9:00am PST 3D cell culture and analysis and the study of organoids and spheroids are becoming more prevalent as a research method in publications as traditional...
    APR 29, 2020 8:00 AM PDT
    C.E. CREDITS
    APR 29, 2020 8:00 AM PDT
    Date: April 29, 2020 Time: 8:00AM PDT, 11:00AM EDT Single cell genomics and other next generation sequencing applications depend strongly on adequate upstream sample preparation. Results can...
    MAR 03, 2020 9:00 AM JST
    C.E. CREDITS
    MAR 03, 2020 9:00 AM JST
    DATE: March 3, 2020 TIME: 9:00am JST A major limitation in the ex vivo expansion of harvested human hematopoietic stem-progenitor cells (HSPCs) is the rapid differentiation of HSPCs at the e...
    Loading Comments...
    Show Resources
    Attendees
    • See more