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Related Concept Videos

MicroRNAs01:22

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
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MicroRNAs01:22

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
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Passenger strand loading in overexpression experiments using microRNA mimics.

Rolf Søkilde1, Inga Newie, Helena Persson

  • 1a Department of Clinical Sciences, Lund ; Division of Oncology and Pathology ; Lund University Cancer Center at Medicon Village ; Lund , Sweden.

RNA Biology
|June 30, 2015
PubMed
Summary

MicroRNA mimics, used in gene function research, can cause unintended side effects. The complementary strand of these mimics may exhibit unwanted activity, impacting experimental accuracy and results.

Keywords:
AGO, ArgonauteArgonaute proteinGEO, Gene Expression OmnibusHITS-CLIP, high-throughput sequencing of RNA isolated by crosslinking immunoprecipitationOAS, oligoadenylate synthetasePKR, protein kinase RRISC, RNA-induced silencing complexRNA interferenceTLR3, Toll-like receptor 3mRNA, mRNAmiRNA, microRNAmicroRNAmimicnt, nucleotideoff-target effectpassenger strandsiRNA

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biotechnology

Background:

  • MicroRNAs (miRNAs) are key regulators of gene expression.
  • Manipulating miRNA function is crucial for basic research.
  • miRNA mimics are synthetic small RNAs designed to imitate endogenous miRNA function.

Purpose of the Study:

  • To investigate potential side effects of miRNA mimics.
  • To assess the accuracy of miRNA mimic-based gain-of-function studies.
  • To identify unintended activities associated with miRNA mimics.

Main Methods:

  • In silico analysis of sequence motifs in microarray expression data.
  • Sequencing of Argonaute (AGO)-associated small RNAs.
  • Evaluation of miRNA mimic complementary strand activity.

Main Results:

  • Analysis revealed over-represented sequence motifs associated with miRNA mimics.
  • Sequencing identified unintended small RNAs derived from miRNA mimics.
  • Evidence suggests the miRNA mimic complementary strand can possess unwanted biological activity.

Conclusions:

  • The use of miRNA mimics may lead to significant off-target effects.
  • Unwanted activity of the miRNA mimic complementary strand can compromise experimental outcomes.
  • Independent validation of miRNA mimic function is recommended before use in research.