Hydrophilic Surface Modification of Cationic Unimolecular Bottlebrush Vectors Moderate pDNA and RNP Bottleplex Stability and Delivery Efficacy

Rishad J. Dalal, Monica L. Ohnsorg, Sidharth Panda, Theresa M. Reineke

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

A cationic unimolecular bottlebrush polymer with chemically modified end-groups was synthesized to understand the impact of hydrophilicity on colloidal stability, nucleic acid delivery performance, and toxicity. The bottlebrush polymer template was synthesized using grafting-through techniques and was therefore composed of a polynorbornene backbone with poly(2-(dimethylamino)ethyl methacrylate) side chains with dodecyl trithiocarbonate end-groups. Postpolymerization modification was performed to fully remove the end-groups or install hydroxy and methoxy poly(ethylene glycol) functional groups on the bottlebrush exterior. The bottlebrush family was preformulated with biological payloads of pDNA and CRISPR-Cas9 RNP in both water and PBS to understand binding, aggregation kinetics, cytotoxicity, and delivery efficacy. Increasing end-group hydrophilicity and preformulation of bottleplexes in PBS increased colloidal stability and cellular viability; however, this did not always result in increased transfection efficiency. The bottlebrush family exemplifies how formulation conditions, polymer loading, and end-group functionality of bottlebrushes can be tuned to balance expression with cytotoxicity ratios and result in enhanced overall performance.

Original languageEnglish (US)
Pages (from-to)5179-5192
Number of pages14
JournalBiomacromolecules
Volume23
Issue number12
DOIs
StatePublished - Dec 12 2022

Bibliographical note

Funding Information:
The authors acknowledge Nanite, Inc. for funding this work. M.L.O. acknowledges funding support from the National Science Foundation Graduate Research Fellowship Program under Grant No. 00039202. The authors acknowledge Craig Van Bruggen for technical advice. The TOC, and A were made by the authors on Biorender.com.

Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.

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