Transcriptional Activation of Human CREB Gene Promoter Using Bis-PNA (Peptide Nucleic Acid)
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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.
KeywordsH293 Cell Peptide Nucleic Acid Template Strand CREB Gene Total CREB
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