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Identification of Immune Modulatory miRNAs by miRNA Enrichment via RNA Affinity Purification

  • Uta Sandy Tretbar
  • Michael Friedrich
  • Maria-Filothei Lazaridou
  • Barbara SeligerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1913)

Abstract

Immune escape by cancer cells can be triggered by aberrant expression of immunological key players, which can be achieved by distinct molecular mechanisms including immune modulatory miRNAs. One suitable method to identify miRNAs that specifically target immune relevant molecules is the miRNA enrichment via RNA affinity purification method named miTRAP (miRNA trapping by RNA in vitro affinity purification). Here, we present a detailed protocol for construct preparation, RNA immobilization via MS2BP-MBP to beads, miRNA enrichment, and elution followed by analysis of the obtained miRNA candidates via qRT-PCR.

Key words

miRNAs RNA in vitro affinity purification miTRAP miRNA enrichment Immune relevant 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Uta Sandy Tretbar
    • 1
  • Michael Friedrich
    • 1
  • Maria-Filothei Lazaridou
    • 1
  • Barbara Seliger
    • 1
    Email author
  1. 1.Institute for Medical ImmunologyMartin-Luther-University Halle-WittenbergHalleGermany

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