Synthesis and characterization of peptide nucleic acid for experimental use of directing anthrapyrazole intercalation

CHED 924

James M. Bradley, bradleyj@southwestern.edu, Chemistry and Biochemistry Department, Southwestern University, 1001 University Ave., Georgetown, TX 78626 and Kerry A. Bruns, brunsk@southwestern.edu, Department of Chemistry, Southwestern University, 1001 E. University Ave., Georgetown, TX 78626.
A peptide nucleic acid (PNA) was designed to be complimentary to the template strand of the viral T7 promoter, and synthesized using Fmoc solid phase peptide synthetic methods. The PNA structure includes two D-lysine residues to enhance DNA binding by imparting two additional positive charges at physiological pH. The PNA was purified by HPLC using a preparative C-18 reversed phase column. The structure of the PNA was verified using MALDI-TOF MS and ESI-MS each with a result of 3470.5g•mol-1. ESI-MS was also used to detect double stranded complexes of the PNA with its target sequence oligonucleotides. Future goals include incorporating an anthrapyrazole (AP) intercalating moiety to the amino terminus of the PNA by way of a multi-ether linker. This may target the intercalator to a selected region of DNA due to the specifity of the PNA/DNA complex formation. Thus, the AP-PNA adducts may display characteristics favorable for prolonged gene-silencing.