Transcriptional activation of human CREB gene promoter using bis-PNA (peptide nucleic acid)

Abstract

Peptide nucleic acid (PNA) is a synthetic nucleotide analogue or mimic consisting of an amino-ethyl backbone with nucleoside bases projecting from amide linkers along the major axis. PNA is able to bind complimentary RNA or DNA through Watson-Crick or Hoogsteen hydrogen bonding. The applications of PNA for antisense or antigene “knock-down” are well-established. In addition, PNA chemistry is unusual in its ability to promote transcription through artificial D-loops when bound to the template strand of DNA in a [PNA]2-DNA conformation. We describe here the application of PNA gene activation chemistry to the human CREB gene, which is involved in diverse neurological processes. A gene-activating bis-PNA targeting the CREB promoter region was annealed to a complimentary DNA carrier oligo and introduced to h293 cells using standard liposomal transfection, and levels of total CREB were quantified on the RNA and protein level from cell lysates.