Absence of unspecific innate immune cell activation by GATA-3-specific DNAzymes

Nucleic Acid Ther. 2012 Apr;22(2):117-26. doi: 10.1089/nat.2011.0294. Epub 2012 Mar 19.


DNAzymes of the 10-23 family represent an important class of antisense molecules with implications for therapeutic treatment of diseases. These molecules are single-stranded oligodeoxynucleotides combining the high specificity of oligonucleotide base pairing with an inherent RNA-cleaving enzymatic activity. However, like other oligonucleotide-based molecules these substances might exert so-called off-target effects, which have not been investigated so far for this molecule class. Therefore, the present study investigates putative off-target effects of DNAzymes on innate immune mechanisms using GATA-3-specific DNAzymes that have recently been developed as novel therapeutic approach for the treatment of allergic diseases including allergic asthma. The conserved catalytic domain of 10-23 DNAzymes contains a CpG motif that may stimulate innate immune cells via Toll-like receptor 9 (TLR-9). Therefore, potential TLR-9-mediated as well as TLR-9 independent cell activation was investigated using TLR-9-transfected HEK293 cells, macrophage cell lines and primary innate immune cells. Furthermore, putative effects of GATA-3-specific DNAzymes on the activation of neutrophil granulocytes and degranulation of mast cells/basophils were analyzed. In summary, no innate immune cell-stimulating activities of the tested DNAzymes were observed in any of the systems. Consequently, use of GATA-3-specific DNAzymes may represent a novel and highly specific approach for the treatment of allergic diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Basophils / drug effects
  • Basophils / metabolism
  • Catalytic Domain
  • Cells, Cultured
  • Cytokines / metabolism
  • DNA, Catalytic / pharmacology*
  • DNA, Single-Stranded / pharmacology*
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism
  • GATA3 Transcription Factor
  • Genes, Reporter
  • Immune System / cytology*
  • Immunity, Innate / drug effects*
  • Immunologic Factors / pharmacology*
  • Luciferases / biosynthesis
  • Luciferases / genetics
  • Macrophages / drug effects
  • Macrophages / immunology
  • Macrophages / metabolism
  • Mast Cells / drug effects
  • Mast Cells / enzymology
  • Mice
  • Mice, Inbred BALB C
  • NF-kappa B / genetics
  • Neutrophils / drug effects
  • Neutrophils / metabolism
  • Primary Cell Culture
  • Rats
  • Respiratory Burst / drug effects
  • Spleen / cytology
  • Toll-Like Receptor 9 / metabolism


  • Cytokines
  • DNA, Catalytic
  • DNA, Single-Stranded
  • GATA3 Transcription Factor
  • Immunologic Factors
  • NF-kappa B
  • RNA-cleaving DNA 10-23
  • Tlr9 protein, mouse
  • Toll-Like Receptor 9
  • Luciferases