Synthesis and applications of oligonucleotide-bound alginate hydrogels

This research investigates the use of circular dichroism (CD) spectroscopy for label-free detection of DNA and RNA hybridization in oligonucleotide-bound alginate hydrogels, with applications in biosensing. Incorporating 2’-fluoro modifications significantly enhanced sensitivity, with 2’-F probes outperforming unmodified DNA and RNA in both detection signal and binding specificity. In mixtures with up to four non-complementary sequences, 2’-F probes also yielded higher classification accuracy using random forest analysis, when integrating CD and thermal melt data. Concurrently, the potential of covalently bound alginate hydrogels for small interfering RNA (siRNA)-mediated gene silencing was evaluated. Covalent attachment resulted in a tenfold decrease in IC50 values compared to unbound siRNA, indicating improved gene silencing efficiency. Additionally, nuclease stability studies revealed that alginate-siRNA conjugates retained double the amount of nondegraded siRNA after 15 minutes in fetal bovine serum compared to unbound siRNA. Together, these findings highlight the versatility of alginate-oligonucleotide platforms in biosensing and therapeutic delivery applications.

Name thesis:degree_name

Master of Science (MSc)

Discipline thesis:degree_discipline

Applied Bioscience

Grantor

University of Ontario Institute of Technology

Year dc:date.issued

2025

Author dc:creator

• Lubrin, Daisee K.

Advisor dc:contributor.advisor

• Stotesbury, Theresa

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/10155/1961

OAI identifier oai:identifier

oai:ontariotechu.scholaris.ca:10155/1961