# Achieving enhanced cytosolic delivery and greater efficacy of therapeutic nucleic acids using DNA-surfactant conjugates

> **NIH NIH R35** · UNIVERSITY OF CONNECTICUT STORRS · 2022 · $402,500

## Abstract

Project Summary
The rapidly expanding field of therapeutic nucleic acids (TNAs) has led to an increased urgency in the
development of chemical approaches that can broaden their clinical application. In just the last 3 years the US
Food and Drug Administration has approved two TNA based drugs that treat diseases that previously had little
option for treatment. Despite these recent achievements, many clinical trials involving TNAs result in termination
due to limited efficacy, a result that is often attributed to poor delivery owing to the highly polar and charged
character of TNAs that limits accessibility to mRNA in the cytosol of cells. Many platforms deliver nucleic acids
using nanoparticle-based approaches that are internalized into cells through endocytosis mechanisms.
Therefore, a central bottleneck to their effective delivery is escaping endosomal compartments and gaining
access to the cytosol. The central vision of my research program is to address these challenges associated with
endosomal escape and TNA delivery to the cytosol at the molecular level. We aim to address these challenges
through the synthesis of nucleic acid surfactant conjugates that we have recently shown can successfully target
mRNA for gene silencing in vitro. As their design is chemically tunable, we aim to systematically assess the role
of the surfactant as it relates to the extent of endosomal escape by evaluating the importance of hydrophobic
character, net charge and size of the conjugates. We will also develop a new class of fluorescent surfactant
probes which can be used to monitored the delivery and stability of the surfactant conjugates as they are
trafficked through cells, thereby helping us to quantify the effects changes in the chemical character of the
conjugates have on cytosolic delivery. In conjunction with these studies we will determine the net effect of
chemically modifying the nucleic acid surfactant conjugates on the efficacy of gene knockdown in vitro using
DNAzymes that result in mRNA cleavage. By controlling the molecular design of individual DNAzyme-surfactant
conjugates we can better understand their mechanism of cellular entry and the properties that lead to cytosolic
access. Through successful realization of our program we will not only contribute to our understanding of the
properties that are necessary for nucleic acids to successfully gain access to the cytosol of cells but also design
a platform that will have immediate therapeutic value suitable for future translational applications.

## Key facts

- **NIH application ID:** 10460258
- **Project number:** 5R35GM138226-03
- **Recipient organization:** UNIVERSITY OF CONNECTICUT STORRS
- **Principal Investigator:** Jessica L Rouge
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $402,500
- **Award type:** 5
- **Project period:** 2020-08-01 → 2025-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10460258

## Citation

> US National Institutes of Health, RePORTER application 10460258, Achieving enhanced cytosolic delivery and greater efficacy of therapeutic nucleic acids using DNA-surfactant conjugates (5R35GM138226-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10460258. Licensed CC0.

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