Lentiviral delivery of RNAi for in vivo lineage-specific modulation of gene expression in mouse lung macrophages

Mol Ther. 2013 Apr;21(4):825-33. doi: 10.1038/mt.2013.19. Epub 2013 Feb 12.


Although RNA interference (RNAi) has become a ubiquitous laboratory tool since its discovery 12 years ago, in vivo delivery to selected cell types remains a major technical challenge. Here, we report the use of lentiviral vectors for long-term in vivo delivery of RNAi selectively to resident alveolar macrophages (AMs), key immune effector cells in the lung. We demonstrate the therapeutic potential of this approach by RNAi-based downregulation of p65 (RelA), a component of the pro-inflammatory transcriptional regulator, nuclear factor κB (NF-κB) and a key participant in lung disease pathogenesis. In vivo RNAi delivery results in decreased induction of NF-κB and downstream neutrophilic chemokines in transduced AMs as well as attenuated lung neutrophilia following stimulation with lipopolysaccharide (LPS). Through concurrent delivery of a novel lentiviral reporter vector (lenti-NF-κB-luc-GFP) we track in vivo expression of NF-κB target genes in real time, a critical step towards extending RNAi-based therapy to longstanding lung diseases. Application of this system reveals that resident AMs persist in the airspaces of mice following the resolution of LPS-induced inflammation, thus allowing these localized cells to be used as effective vehicles for prolonged RNAi delivery in disease settings.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cells, Cultured
  • Gene Expression Regulation / genetics
  • Gene Expression Regulation / physiology
  • Lentivirus / genetics*
  • Lung / immunology*
  • Lung / metabolism*
  • Macrophages / metabolism*
  • Macrophages, Alveolar / metabolism*
  • Mice
  • NF-kappa B / genetics
  • RNA Interference / physiology
  • Transcription Factor RelA / genetics


  • NF-kappa B
  • Transcription Factor RelA