Skip to main content

RADPRE: A Computational Program for Identification of Differential mRNA Processing Including Alternative Polyadenylation

  • Protocol
  • First Online:
Polyadenylation in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1255))

Abstract

Genome-wide studies revealed the prevalence of multiple transcripts resulting from alternative polyadenylation (APA) of a single given gene in higher eukaryotes. Several studies in the past few years attempted to address how those APA events are regulated and what the biological consequences of those regulations are. Common to these efforts is the comparison of unbiased transcriptome data, either derived from whole-genome tiling array or next generation sequencing, to identify the specific APA events in a given condition. RADPRE (Ratio-based Analysis of Differential mRNA Processing and Expression) is an R program, developed to serve such a purpose using data from the whole-genome tilling array. RADPRE took a set of tilling array data as input, performed a series of calculation including a correction of the probe affinity variation, a hierarchy of statistical tests and an estimation of the false discovery rate (FDR) of the differentially processed genes (DPG). The result was an output of a few tabular files including DPG and their corresponding FDR. This chapter is written for scientists with limited programming experiences.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Jan CH, Friedman RC, Ruby JG, Bartel DP (2011) Formation, regulation and evolution of Caenorhabditis elegans 3′UTRs. Nature 469(7328):97–101. doi:10.1038/nature09616

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Wu X, Liu M, Downie B, Liang C, Ji G, Li QQ, Hunt AG (2011) Genome-wide landscape of polyadenylation in Arabidopsis provides evidence for extensive alternative polyadenylation. Proc Natl Acad Sci U S A 108(30):12533–12538. doi:10.1073/pnas.1019732108

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Marquez Y, Brown JW, Simpson C, Barta A, Kalyna M (2012) Transcriptome survey reveals increased complexity of the alternative splicing landscape in Arabidopsis. Genome Res 22(6):1184–1195. doi:10.1101/gr.134106.111

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Thomas PE, Wu X, Liu M, Gaffney B, Ji G, Li QQ, Hunt AG (2012) Genome-wide control of polyadenylation site choice by CPSF30 in Arabidopsis. Plant Cell 24(11):4376–4388. doi:10.1105/tpc.112.096107

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Zheng J, Xing D, Wu X, Shen Y, Kroll DM, Ji G, Li QQ (2011) Ratio-based analysis of differential mRNA processing and expression of a polyadenylation factor mutant pcfs4 using arabidopsis tiling microarray. PLoS One 6(2):e14719. doi:10.1371/journal.pone.0014719

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Huber W, von Heydebreck A, Sultmann H, Poustka A, Vingron M (2002) Variance stabilization applied to microarray data calibration and to the quantification of differential expression. Bioinformatics 18(Suppl 1):S96–S104

    Article  PubMed  Google Scholar 

  7. Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U, Speed TP (2003) Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4(2):249–264. doi:10.1093/biostatistics/4.2.249

    Article  PubMed  Google Scholar 

  8. Tusher VG, Tibshirani R, Chu G (2001) Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 98(9):5116–5121. doi:10.1073/pnas.091062498

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Jones-Rhoades MW, Borevitz JO, Preuss D (2007) Genome-wide expression profiling of the Arabidopsis female gametophyte identifies families of small, secreted proteins. PLoS Genet 3(10):1848–1861. doi: 10.1371/journal.pgen.0030171

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgement

The authors appreciate the original contributions of Jianti Zheng and Guoli Ji. The project was supported by grants from US National Science Foundation (grant nos. IOS–0817829 and IOS-1353354 to QQL), and from Ohio Plant Biotech Consortium (to QQL and DX).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Denghui Xing .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Xing, D., Li, Q.Q. (2015). RADPRE: A Computational Program for Identification of Differential mRNA Processing Including Alternative Polyadenylation. In: Hunt, A., Li, Q. (eds) Polyadenylation in Plants. Methods in Molecular Biology, vol 1255. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2175-1_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-2175-1_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2174-4

  • Online ISBN: 978-1-4939-2175-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics