Reverse-transcriptase polymerase chain reaction to detect extracellular mRNAs

Sweta Rani, Lorraine O'Driscoll

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Citations (Scopus)

Abstract

The presence of extracellular nucleic acids has been reported in serum/plasma from cancer and diabetes patients that may help in disease diagnosis. Taking insulin-producing cells as examples here, RT-PCR was used to investigate a correlation between the presence and amounts of extracellular mRNA(s) and cell mass and/or function. RT-PCR was performed on a range of mRNAs, including Pdx1, Npy, Egr1, Pld1, Chgb, InsI, InsII, and Actb in biological triplicate analyses. Reproducible amplification of these mRNAs from MIN6, MIN6 B1, and Vero-PPI cells and their CM suggests that beta cells transcribe and release these mRNAs into their environment. mRNAs secreted from insulin-producing cells into their extracellular environment may have potential as extracellular biomarkers for assessing beta cell mass and function.

Original languageEnglish
Title of host publicationGene Expression Profiling
Subtitle of host publicationMethods and Protocols
EditorsLorraine O'Driscoll
Pages15-25
Number of pages11
DOIs
Publication statusPublished - 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume784
ISSN (Print)1064-3745

Keywords

  • Beta cell function
  • Beta cell mass
  • Conditioned media
  • Extracellular nucleic acid
  • Insulin-producing cells
  • MIN6
  • RT-PCR

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