title: Anti-gene padlocks eliminate Escherichia coli based on their genotype.
abstract: OBJECTIVE
  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 a double 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 approximately 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.
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA.
authors.forename: Chanjuan
authors.initials: C
authors.lastname: Shi
authors.forename: Antony R
authors.initials: AR
authors.lastname: Parker
authors.forename: Li
authors.initials: L
authors.lastname: Hua
authors.forename: Craig N
authors.initials: CN
authors.lastname: Morrell
authors.forename: Soo Chin
authors.initials: SC
authors.lastname: Lee
authors.forename: Viswanath
authors.initials: V
authors.lastname: Bandaru
authors.forename: J Stephen
authors.initials: JS
authors.lastname: Dumler
authors.forename: T C
authors.initials: TC
authors.lastname: Wu
authors.forename: James R
authors.initials: JR
authors.lastname: Eshleman
mesh_terms: D004926:Escherichia coli / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods / Q000639:trends*; D005838:Genotype; D015335:Molecular Probes / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics
chemical_list: D015335:Molecular Probes; D009841:Oligonucleotides