Automated Fast-Flow Synthesis of Peptide Nucleic Acid Conjugates

24 February 2021, Version 1


Peptide nucleic acids (PNAs) are charge-neutral oligonucleotides with emerging potential for treatment of genetic, acquired, and viral diseases, including COVID-19. Their challenging synthesis, however, limits their use for rapid therapeutic intervention and widespread application. Here, we report a highly efficient technology that utilizes a fully automated fast-flow instrument to manufacture cell-penetrating peptide-conjugated PNAs (PPNAs) in a single shot. The machine is rapid: each amide bond is formed in 10 seconds and the synthesis of an 18-mer bioactive PNA is complete in one hour. Anti-IVS2-654 PPNA synthesized in a single shot with this instrument presented over 16-fold activity compared to unmodified PNA in a splice-correction assay. We demonstrated the utility of this approach by chemically synthesizing an eight-member anti-SARS-CoV-2 PPNA library within one day. A designer PPNA targeting the 5’ untranslated region of SARS-CoV-2 genomic RNA reduced the viral titer by over 95% in live virus infection assays (IC50: 0.8 mM). Our technology can rapidly yield on-demand PPNA candidates to tackle newly-emerging viral pathogens.


Peptide nucleic acid
Flow chemistry
antisense therapeutics
process Optimization
Oligonucleotides Conjugated

Supplementary materials



Comments are not moderated before they are posted, but they can be removed by the site moderators if they are found to be in contravention of our Commenting Policy [opens in a new tab] - please read this policy before you post. Comments should be used for scholarly discussion of the content in question. You can find more information about how to use the commenting feature here [opens in a new tab] .
This site is protected by reCAPTCHA and the Google Privacy Policy [opens in a new tab] and Terms of Service [opens in a new tab] apply.