Skip to main content
SpringerLink
Log in

Analysis of RBP Regulation and Co-regulation of mRNA 3′ UTR Regions in a Luciferase Reporter System

  • Protocol
  • First Online:
RNA Abundance Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2170))

Abstract

The luciferase reporter assay is a widely used tool to study the cis and trans factors controlling regulation of gene expression. In this assay, regulatory elements can be fused to the luciferase gene, and as a result effect protein output by changing rates of transcription, rates of translation, or mRNA stability. This protocol focuses on probing the function of RNA-binding proteins (RBPs) through their interactions with the 3′ untranslated region (UTR), thus examining gene expression regulation on the mRNA level. Assessment of 3′ UTR sequence requirements, as well as single and co-regulatory roles of RBPs in regulation of mRNAs will be discussed.

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

Access this chapter

Log in via an institution
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 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Castello A, Fischer B, Eichelbaum K, Horos R, Beckmann BM, Strein C, Davey NE, Humphreys DT, Preiss T, Steinmetz LM, Krijgsveld J, Hentze MW (2012) Insights into RNA biology from an atlas of mammalian mRNA-binding proteins. Cell 149:1393

    Article  CAS  Google Scholar 

  2. Hentze MW, Castello A, Schwarzl T, Preiss T (2018) A brave new world of RNA-binding proteins. Nat Rev Mol Cell Biol 19:327

    Article  CAS  Google Scholar 

  3. Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136:215

    Article  CAS  Google Scholar 

  4. Lunde BM, Moore C, Varani G (2007) RNA-binding proteins: modular design for efficient function. Nat Rev Mol Cell Biol 8:479

    Article  CAS  Google Scholar 

  5. Li Y, Estep JA, Karginov FV (2018) Transcriptome-wide identification and validation of interactions between the miRNA machinery and HuR on mRNA targets. J Mol Biol 430:285

    Article  CAS  Google Scholar 

  6. Kedde M, van Kouwenhove M, Zwart W, Oude Vrielink JA, Elkon R, Agami R (2010) A Pumilio-induced RNA structure switch in p27-3' UTR controls miR-221 and miR-222 accessibility. Nat Cell Biol 12:1014

    Article  CAS  Google Scholar 

  7. Weidmann CA, Goldstrohm AC (2012) Drosophila pumilio protein contains multiple autonomous repression domains that regulate mRNAs independently of Nanos and brain tumor. Mol Cell Biol 32:527

    Article  CAS  Google Scholar 

  8. Sherf B, Navarro S, Hannah R, Wood K (1997) Co-reporter technology integrating firefly and Renilla luciferase assays. In: Hastings JW, Kricka LJ, Stanley PE (eds) 9th international symposium on bioluminescence and Chemiluminescence. John Wiley & Suns Ltd., Chichester, pp 228–231

    Google Scholar 

  9. Sanjana NE, Cong L, Zhou Y, Cunniff MM, Feng G, Zhang F (2012) A transcription activator-like effector toolbox for genome engineering. Nat Protoc 7:171

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Molecular, Cell and Systems Biology, Institute for Integrative Genome Biology, University of California, Riverside, CA, USA

    Erin L. Sternburg & Fedor V. Karginov

Authors
  1. Erin L. Sternburg
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fedor V. Karginov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fedor V. Karginov .

Editor information

Editors and Affiliations

  1. Department of Plant Pathology and Microbiology, Center for Plant Cell Biology and Institute for Integrative Genome Biology, University of California, Riverside, CA, USA

    Hailing Jin

  2. Department of Nematology, University of California, Riverside, CA, USA

    Isgouhi Kaloshian

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Sternburg, E.L., Karginov, F.V. (2021). Analysis of RBP Regulation and Co-regulation of mRNA 3′ UTR Regions in a Luciferase Reporter System. In: Jin, H., Kaloshian, I. (eds) RNA Abundance Analysis . Methods in Molecular Biology, vol 2170. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0743-5_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0743-5_7

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0742-8

  • Online ISBN: 978-1-0716-0743-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation