Our research on Peptide Nucleic Acids (abbreviated as PNAs) focuses on introducing chemical modifications that will make this class of molecules broadly useful to detect sequences of DNA. Unique DNA sequences are associated with diseases, pathogens, and many agents associated with bioterrorism. Detection of DNA from these agents can be employed as a method to detect their presence or absence. Our research involves the synthesis of a class of non-natural molecules (called PNAs) that bind to specific DNA sequences. We can design our molecules to bind to any sequence of DNA, and previously we have found that our molecules are extremely good at selective recognition of DNA associated with anthrax. During the past year, we have imporved the synthesis of the building blocks we need to make selective PNAs that bind to anthrax DNA, and we have developed, and published, a color-based detection assay using our PNA molecules to detect anthrax DNA. We have also used real DNA isolated from the anthrax bacteria to comfirm that our methods can work with a real system. In addition, we have coupled our methods with fluorescent groups to create light-up probes that will only fluoresce in the presence of target DNA. PNAs are also useful as antisense and antigene molecules, however delivery into cells has been difficult. This past year we have begun a collaboration with researchers in the NCI and FDA examining the delivery of PNA to cells using an SV40 delivery system.

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2007
Total Cost
$439,398
Indirect Cost
City
State
Country
United States
Zip Code
Zhang, Ning; Appella, Daniel H (2010) Advantages of peptide nucleic acids as diagnostic platforms for detection of nucleic acids in resource-limited settings. J Infect Dis 201 Suppl 1:S42-5
Englund, Ethan A; Appella, Daniel H (2007) Gamma-substituted peptide nucleic acids constructed from L-lysine are a versatile scaffold for multifunctional display. Angew Chem Int Ed Engl 46:1414-8
Zhang, Ning; Appella, Daniel H (2007) Colorimetric detection of anthrax DNA with a Peptide nucleic acid sandwich-hybridization assay. J Am Chem Soc 129:8424-5
Witschi, Mark A; Pokorski, Jonathan K; Appella, Daniel H (2006) Peptide nucleic acid microarrays made with (S,S)-trans-cyclopentane-constrained peptide nucleic acids. Methods Enzymol 410:189-200
Englund, Ethan A; Xu, Qun; Witschi, Mark A et al. (2006) PNA-DNA duplexes, triplexes, and quadruplexes are stabilized with trans-cyclopentane units. J Am Chem Soc 128:16456-7
Xu, Qun; Appella, Daniel H (2006) Practical synthesis of trans-tert-butyl-2-aminocyclopentylcarbamate and resolution of enantiomers. J Org Chem 71:8655-7
Pokorski, Jonathan K; Nam, Jwa-Min; Vega, Rafael A et al. (2005) Cyclopentane-modified PNA improves the sensitivity of nanoparticle-based scanometric DNA detection. Chem Commun (Camb) :2101-3
Guler, Mustafa O; Pokorski, Jonathan K; Appella, Daniel H et al. (2005) Enhanced oligonucleotide binding to self-assembled nanofibers. Bioconjug Chem 16:501-3
Englund, Ethan A; Appella, Daniel H (2005) Synthesis of gamma-substituted peptide nucleic acids: a new place to attach fluorophores without affecting DNA binding. Org Lett 7:3465-7