Anti-gene padlocks eliminate Escherichia coli based on their genotype

Chanjuan Shi, Antony R. Parker, Li Hua, Craig N. Morrell, Soo Chin Lee, Viswanath Bandaru, J. Stephen Dumler, T. C. Wu, James R. Eshleman

Research output: Contribution to journalArticle

Abstract

Objectives: Several therapeutic strategies that target nucleic acids exist; however, most approaches target messenger RNA, rather than genomic DNA. We describe a novel oligonucleotide-based strategy, called anti-gene padlocks (AGPs), which eliminate Escherichia coli based on their genotype. Methods: The strategy employs an oligonucleotide with adouble hairpin structure where both strands of the AGP are complementary to both strands of a target gene. We tested AGPs for in vitro binding and inhibition of DNA polymerization. AGPs were electroporated into bacterial cells with and without gene targets along with an ampicillin resistance plasmid, and cell survival was measured. Results: In vitro, AGPs bound the DNA target in a sequence-dependent fashion and inhibited DNA synthesis. When transformed into bacterial cells containing 10, 20 or 30 bp lacZ or 20 bp proA DNA targets in their genomes, AGPs selectively killed or otherwise inhibited growth of these cells, while those lacking the target demonstrated little, if any, toxicity. A single transformation resulted in ∼30% to 40% loss of target-bearing cells. Structure-function experiments were performed to define essential AGP requirements. Conclusions: These results suggest that AGPs may be a useful tool to eliminate specific cell populations.

Original languageEnglish (US)
Pages (from-to)262-272
Number of pages11
JournalJournal of Antimicrobial Chemotherapy
Volume61
Issue number2
DOIs
StatePublished - Feb 1 2008

Keywords

  • Antibiotics
  • Antimicrobial agents
  • Gene targeting
  • Novel therapeutics
  • Oligonucleotide
  • Padlock probes

ASJC Scopus subject areas

  • Pharmacology
  • Microbiology (medical)
  • Infectious Diseases
  • Pharmacology (medical)

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  • Cite this

    Shi, C., Parker, A. R., Hua, L., Morrell, C. N., Lee, S. C., Bandaru, V., Dumler, J. S., Wu, T. C., & Eshleman, J. R. (2008). Anti-gene padlocks eliminate Escherichia coli based on their genotype. Journal of Antimicrobial Chemotherapy, 61(2), 262-272. https://doi.org/10.1093/jac/dkm482