DNA Aptamer Induction of Anthrax Germination

OmniSite BioDiagnostics Inc. proposes to determine the feasibility of using DNA Aptamers to bind key germinant receptors on nonpathogenic Sterne strain anthrax spores to induce synchronous germination, hence leading to more effective antibiotic treatmentof inhalation anthrax. DNA Aptamers are short oligonucleotides that act like antibodies by binding any target against which they are raised with very high affinity and specificity. Aptamers are generated by a process called SELEX (Systematic Evolution ofLigands by EXponential enrichment) consisting of alternating iterations of affinity selection (from a random DNA library allowed to interact with the immobilized target), washing and PCR amplification of selected DNA sequences that bind the target. Dr.Bruno (proposed PI and CTO of OmniSite) is well versed and published in the area of Aptamer generation and assay development. In fact, Dr. Bruno made and published on the first anthrax spore binding DNA Aptamers (Bruno & Kiel, Biosensors & Bioelectronics1999). OmniSite proposes separation and characterization of inner and outer cortex proteins (by PAGE followed by Coomassie and silver staining) from anthrax spores with generation of aptamers to each identified protein. Then OmniSite will test theaptamers generated for their ability to enhance or inhibit spore germination by colony counts. This proposed project has several major scientific benefits in that, if successful, it may uncover previously unknown spore receptors that turn on germination and it will increase scientific understanding of the germination process. In a business sense,the project may lead to new therapeutic Aptamers capable of enhancing spore germination so that ciprofloxacin and other antibiotics can be more effective at killing the vegetative form of B. anthracis. In a broader sense too, technology gained byperformance of the project may lead to new Aptamer-based ways to kill or inhibit pathogenic bacteria.