Next-generation sequencing identifies the natural killer cell microRNA transcriptome

Genome Res. 2010 Nov;20(11):1590-604. doi: 10.1101/gr.107995.110. Epub 2010 Oct 8.


Natural killer (NK) cells are innate lymphocytes important for early host defense against infectious pathogens and surveillance against malignant transformation. Resting murine NK cells regulate the translation of effector molecule mRNAs (e.g., granzyme B, GzmB) through unclear molecular mechanisms. MicroRNAs (miRNAs) are small noncoding RNAs that post-transcriptionally regulate the translation of their mRNA targets, and are therefore candidates for mediating this control process. While the expression and importance of miRNAs in T and B lymphocytes have been established, little is known about miRNAs in NK cells. Here, we used two next-generation sequencing (NGS) platforms to define the miRNA transcriptomes of resting and cytokine-activated primary murine NK cells, with confirmation by quantitative real-time PCR (qRT-PCR) and microarrays. We delineate a bioinformatics analysis pipeline that identified 302 known and 21 novel mature miRNAs from sequences obtained from NK cell small RNA libraries. These miRNAs are expressed over a broad range and exhibit isomiR complexity, and a subset is differentially expressed following cytokine activation. Using these miRNA NGS data, miR-223 was identified as a mature miRNA present in resting NK cells with decreased expression following cytokine activation. Furthermore, we demonstrate that miR-223 specifically targets the 3' untranslated region of murine GzmB in vitro, indicating that this miRNA may contribute to control of GzmB translation in resting NK cells. Thus, the sequenced NK cell miRNA transcriptome provides a valuable framework for further elucidation of miRNA expression and function in NK cell biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Animals
  • Base Sequence
  • Cells, Cultured
  • Computational Biology / instrumentation
  • Computational Biology / methods
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation / drug effects
  • Granzymes / genetics
  • High-Throughput Nucleotide Sequencing / instrumentation
  • High-Throughput Nucleotide Sequencing / methods*
  • Interleukin-15 / pharmacology
  • Killer Cells, Natural / metabolism*
  • Lymphocyte Activation / drug effects
  • Lymphocyte Activation / genetics
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / genetics*
  • MicroRNAs / isolation & purification
  • MicroRNAs / metabolism
  • MicroRNAs / physiology
  • Molecular Sequence Data
  • Nucleic Acid Hybridization / methods
  • Sequence Analysis, RNA / instrumentation
  • Sequence Analysis, RNA / methods
  • Sequence Homology, Nucleic Acid


  • Interleukin-15
  • MIRN223 microRNA, mouse
  • MicroRNAs
  • Granzymes
  • Gzmb protein, mouse

Associated data

  • GEO/GSE21003