Synthesis and Characterization of Click Nucleic Acid Conjugated Polymeric Microparticles for DNA Delivery Applications
journal contributionposted on 2021-02-23, 17:08 authored by Alex J. Anderson, Emerson Grey, Nicholas J. Bongiardina, Christopher N. Bowman, Stephanie J. Bryant
Microparticle-mediated nucleic acid delivery is a popular strategy to achieve therapeutic outcomes via antisense gene therapy. However, current methods used to fabricate polymeric microparticles suffer from suboptimal properties such as particle polydispersity and low encapsulation efficiency. Here, a new particulate delivery system based on step-growth thiol-Michael dispersion polymerization is reported in which a low polydispersity microparticle is functionalized with a synthetic nucleic acid mimic, namely, click nucleic acids (CNA). CNA oligomers, exhibiting an average length of approximately four nucleic acid repeat units per chain for both adenine and thymine bases, were successfully conjugated to excess thiols present in the microparticles. Effective DNA loading was obtained by simple mixing, and up to 6 ± 2 pmol of complementary DNA/mg of particle was achieved, depending on the length of DNA used. In addition, DNA loading was orders of magnitude less for noncomplementary sequences and sequences containing an alternating base mismatch. The DNA release properties were evaluated, and it was found that release could be triggered by sudden changes in temperature but was unaffected over a range of pH. Finally, phagocytosis of loaded microparticles was observed by confocal microscopy and corroborated by an increase in cellular metabolic activity up to 90%. Overall, this work suggests that CNA functionalized microparticles could be a promising platform for controlled DNA delivery.
Click Nucleicthymine basesCNA functionalized microparticlespolydispersity microparticleDNA loadingConjugated Polymeric Microparticlesparticle polydispersityDNA release propertiesstep-growth thiol-Michael dispersio...DNA deliveryEffective DNA loadingdelivery systemacid deliveryDNA Delivery Applications Micropart...base mismatchencapsulation efficiencynoncomplementary sequencesconfocal microscopyCNA oligomersantisense gene therapysuboptimal properties