Skip to main content

Multiplexed Detection and Quantitation of Extracellular Vesicle RNA Expression Using NanoString

  • Protocol
Extracellular RNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1740))

Abstract

Several different types of RNA molecules such as microRNAs (miRNAs) have been detected within extracellular vesicles in the circulation. The detection and potential utility of these as disease biomarkers requires the ability to detect their presence with adequate sensitivity and to quantitate their expression. The potential for circulating miRNA to serve as biomarkers can be evaluated through their detection in association with specific disease states. Multiplexed detection of several miRNA simultaneously can be useful for discovery studies. We describe the analysis of miRNA from biological fluids like plasma and serum using the Nanostring nCounter platform. Assays can be used to quantitate the expression of miRNA using direct detection based on hybridization to target specific color-coded probes followed by counting each color-coded barcode digitally.

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 69.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 89.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 119.99
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. Tsang HF, Xue VW, Koh SP, Chiu YM, Ng LP, Wong SC (2017) NanoString, a novel digital color-coded barcode technology: current and future applications in molecular diagnostics. Expert Rev Mol Diagn 17(1):95–103. https://doi.org/10.1080/14737159.2017.1268533

    Article  CAS  PubMed  Google Scholar 

  2. Geiss GK, Bumgarner RE, Birditt B, Dahl T, Dowidar N, Dunaway DL, Fell HP, Ferree S, George RD, Grogan T, James JJ, Maysuria M, Mitton JD, Oliveri P, Osborn JL, Peng T, Ratcliffe AL, Webster PJ, Davidson EH, Hood L, Dimitrov K (2008) Direct multiplexed measurement of gene expression with color-coded probe pairs. Nat Biotechnol 26(3):317–325. https://doi.org/10.1038/nbt1385

    Article  CAS  PubMed  Google Scholar 

  3. Markowitz J, Abrams Z, Jacob NK, Zhang X, Hassani JN, Latchana N, Wei L, Regan KE, Brooks TR, Uppati SR, Levine KM, Bekaii-Saab T, Kendra KL, Lesinski GB, Howard JH, Olencki T, Payne PR, Carson WE 3rd (2016) MicroRNA profiling of patient plasma for clinical trials using bioinformatics and biostatistical approaches. Onco Targets Ther 9:5931–5941. https://doi.org/10.2147/OTT.S106288

    Article  PubMed  PubMed Central  Google Scholar 

  4. Das K, Chan XB, Epstein D, Teh BT, Kim KM, Kim ST, Park SH, Kang WK, Rozen S, Lee J, Tan P (2016) NanoString expression profiling identifies candidate biomarkers of RAD001 response in metastatic gastric cancer. ESMO Open 1(1):e000009. https://doi.org/10.1136/esmoopen-2015-000009

    Article  PubMed  PubMed Central  Google Scholar 

  5. Oikonomopoulos A, Polytarchou C, Joshi S, Hommes DW, Iliopoulos D (2016) Identification of circulating MicroRNA signatures in Crohn’s disease using the Nanostring nCounter technology. Inflamm Bowel Dis 22(9):2063–2069. https://doi.org/10.1097/MIB.0000000000000883

    Article  PubMed  Google Scholar 

  6. Wang H, Horbinski C, Wu H, Liu Y, Sheng S, Liu J, Weiss H, Stromberg AJ, Wang C (2016) NanoStringDiff: a novel statistical method for differential expression analysis based on NanoString nCounter data. Nucleic Acids Res 44(20):e151. https://doi.org/10.1093/nar/gkw677

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Veldman-Jones MH, Brant R, Rooney C, Geh C, Emery H, Harbron CG, Wappett M, Sharpe A, Dymond M, Barrett JC, Harrington EA, Marshall G (2015) Evaluating robustness and sensitivity of the NanoString technologies nCounter platform to enable multiplexed gene expression analysis of clinical samples. Cancer Res 75(13):2587–2593. https://doi.org/10.1158/0008-5472.CAN-15-0262

    Article  CAS  PubMed  Google Scholar 

  8. Jung SH, Sohn I (2014) Statistical issues in the design and analysis of nCounter projects. Cancer Inform 13(Suppl 7):35–43. https://doi.org/10.4137/CIN.S16343

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by the Office of the Director, National Institutes of Health (USA) through award UH3 TR000884. We acknowledge the expert assistance of Caitlyn Foerst and thank the members of our laboratories for their contributions.

Disclosures: None.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tushar Patel .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Shukla, N., Yan, I.K., Patel, T. (2018). Multiplexed Detection and Quantitation of Extracellular Vesicle RNA Expression Using NanoString. In: Patel, T. (eds) Extracellular RNA. Methods in Molecular Biology, vol 1740. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7652-2_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7652-2_14

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7651-5

  • Online ISBN: 978-1-4939-7652-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics