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RT-qPCR Detection of Senescence-Associated Circular RNAs

  • Amaresh C. Panda
  • Kotb AbdelmohsenEmail author
  • Myriam Gorospe
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1534)

Abstract

Primary cells that reach the end of their replicative potential, encounter sublethal stress, or experience the activation of certain oncogenes cease proliferation and enter a state of long-term growth arrest named senescence. The senescent process has been implicated in a variety of age-related diseases and also in the pathogenesis of cancer. Senescence is characterized by distinct changes in the types and levels of coding RNAs (mRNAs) as well as in the vast collective of regulatory noncoding (nc)RNAs, which includes microRNAs, long noncoding RNAs (lncRNAs), and circular (circRNAs). Numerous technologies permit the detection of senescence-associated linear transcripts (mRNAs, lncRNAs, microRNAs), but the identification and quantification of circRNAs in senescence require distinct molecular approaches. In this chapter, we describe a method for the detection and measurement of circRNAs in senescent cells using specialized reverse transcription (RT) followed by real-time quantitative (q)PCR analysis.

Key words

RNA-binding proteins Sponge circRNAs Divergent primer design Transcriptome 

Notes

Acknowledgments

This work was supported in its entirety by the National Institute on Aging Intramural Research Program of the National Institutes of Health.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Amaresh C. Panda
    • 1
  • Kotb Abdelmohsen
    • 1
    Email author
  • Myriam Gorospe
    • 1
  1. 1.Laboratory of Genetics and GenomicsNational Institute on Aging-Intramural Research Program, National Institutes of HealthBaltimoreUSA

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