Dendrimer-enabled DNA delivery and transformation of Chlamydia pneumoniae

Hervé C. Gérard, Manoj K. Mishra, Guangzhao Mao, Sunxi Wang, Mirabela Hali, Judith A. Whittum-Hudson, Rangaramanujam M. Kannan, Alan P. Hudson

Research output: Contribution to journalArticlepeer-review

Abstract

The chlamydiae are important human pathogens. Lack of a genetic manipulation system has impeded understanding of the molecular bases of virulence for these bacteria. We developed a dendrimer-enabled system for transformation of chlamydiae and used it to characterize the effects of inserting the C. trachomatis plasmid into C. pneumoniae, which lacks any plasmids. The plasmid was cloned into modified yeast vector pEG(KG) and the clone complexed to polyamidoamine dendrimers, producing 50-100. nm spherical particles. HEp-2 cell cultures were infected with C. pneumoniae strain AR-39. Twenty-four hours later, medium was replaced for 3. hours with dendrimer-plasmid complexes, then removed and the medium replaced. Cultures were harvested at various times post-transformation. Real-time PCR and RT-PCR of nucleic acids from transformed cultures demonstrated plasmid replication and gene expression. The cloned plasmid was replicated and expressed in transformants over 5 passages. This system will allow study of chlamydial gene function, allowing development of novel dendrimer-based therapies. From the Clinical Editor: This team of investigators developed a dendrimer-enabled system for transformation of chlamydiae and successfully utilized it to characterize the effects of inserting the C. trachomatis plasmid into C. pneumonia. This system will allow study of chlamydial gene function, allowing development of novel dendrimer-based therapies.

Original languageEnglish (US)
Pages (from-to)996-1008
Number of pages13
JournalNanomedicine: Nanotechnology, Biology, and Medicine
Volume9
Issue number7
DOIs
StatePublished - Oct 2013

Keywords

  • Chlamydia
  • Genetics
  • Pathogenesis
  • Transcription
  • Transformation

ASJC Scopus subject areas

  • Bioengineering
  • Medicine (miscellaneous)
  • Molecular Medicine
  • Biomedical Engineering
  • Materials Science(all)
  • Pharmaceutical Science

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