Anti-gene padlocks eliminate Escherichia coli based on their genotype.

Published

Journal Article

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 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.

Full Text

Duke Authors

Cited Authors

  • Shi, C; Parker, AR; Hua, L; Morrell, CN; Lee, SC; Bandaru, V; Dumler, JS; Wu, TC; Eshleman, JR

Published Date

  • February 2008

Published In

Volume / Issue

  • 61 / 2

Start / End Page

  • 262 - 272

PubMed ID

  • 18156610

Pubmed Central ID

  • 18156610

Electronic International Standard Serial Number (EISSN)

  • 1460-2091

Digital Object Identifier (DOI)

  • 10.1093/jac/dkm482

Language

  • eng

Conference Location

  • England