Submission Date

4-26-2024

Document Type

Paper- Restricted to Campus Access

Department

Biochemistry & Molecular Biology

Adviser

Samantha Wilner

Committee Member

Amanda J. Reig

Committee Member

Erica Gorenberg

Department Chair

Eric Williamsen

Department Chair

Anthony Lobo

Project Description

The discovery of new drugs often accompanies the need for improved drug delivery vehicles that tackle common problems associated with drug solubility, stability, and biodistribution. Janus dendrimers (JDs), characterized by their branched amphipathic structure, self-assemble into vesicles known as dendrimersomes, which have potential to address these challenges. Notably, Janus dendrimersomes are biocompatible, exhibit mechanical stability, and can encapsulate both hydrophobic and hydrophilic therapeutic cargo. This work seeks to expand the versatility of Janus dendrimersomes in drug delivery in two ways, namely by developing a cell-targeted delivery system using nucleic acid aptamers and by exploring encapsulation and delivery of nucleic acid therapeutics, specifically small interfering RNA (siRNA). First, using azido-modified JDs, we show that DNA-conjugated dendrimers (JD-DNAs) can be synthesized in a one-step reaction with alkyne-modified DNA using copper catalyzed azide-alkyne cycloaddition. These JD-DNAs assemble into dendrimersomes via thin film hydration and can be delivered to cells in a targeted manner via aptamers, or nucleic acid binding species, that bind to the human transferrin receptor. Second, we investigate the ability of ionizable amphiphilic JDs (IAJDs) to form dendrimersome nanoparticles (DNPs) and encapsulate RNA cargo. Specifically, we explore how assembly method, buffer conditions, and presence of polyadenylated (polyA) mRNA affect DNP size and polydispersity index. We also begin to develop a platform for using these DNPs in small interfering RNA (siRNA) delivery, first by creating a cell line that transiently expresses enhanced green fluorescent protein (eGFP). Knockdown of eGFP expression using DNPs encapsulating anti-eGFP siRNA will later serve as a proof-of-concept approach to screen DNPs in siRNA delivery. Taken together, this work showcases new approaches for utilizing JDs in drug delivery applications.

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