Nuclear-targeted CRISPR Delivery By Using Aptamer-modified Polymer Lipid Nanoparticles

Xu, Zichen

Nuclear-targeted CRISPR Delivery By Using Aptamer-modified Polymer Lipid Nanoparticles

Xu, Zichen

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Publication Type:: Thesis
Issue Date:: 2025

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Field	Value	Language
dc.contributor.author	Xu, Zichen
dc.date.accessioned	2026-10-14T03:07:24Z
dc.date.available	2026-10-14T03:07:24Z
dc.date.issued	2025
dc.identifier.uri	http://hdl.handle.net/10453/197221
dc.description	University of Technology Sydney. Faculty of Engineering and Information Technology.	en_US.UTF-8
dc.description.abstract	Efficient clinical translation of CRISPR/Cas9 gene editing requires delivery platforms capable of overcoming intracellular barriers and promoting nuclear localization. In this study, aptamer-functionalized polymer-lipid hybrid nanoparticles (PLHNPs) were developed as a nuclear-targeted non-viral delivery system for CRISPR/Cas9 in cancer cells. The nanoparticles consisted of a biodegradable PLGA core and a lipid shell conjugated with the AS1411 aptamer, which targets nucleolin overexpressed on the membrane and in the nuclei of many cancer cells. This design was intended to enhance both cellular uptake and nuclear transport. The PLHNPs were prepared by nanoprecipitation and optimized for size, polydispersity, surface charge, and loading efficiency. Aptamer conjugation was confirmed by FTIR and zeta potential analysis. In MDA-MB-231 and HCT-116 cells, AS1411-modified PLHNPs showed enhanced uptake and effective nuclear accumulation of CRISPR/Cas9 complexes, as demonstrated by 3D confocal imaging. Functional studies targeting Lcn2 showed increased indel formation, reduced gene expression, and suppressed cell migration. Compared with non-targeted systems, the aptamer-modified PLHNPs achieved approximately 30% higher editing efficiency.	en_US.UTF-8
dc.format	Thesis (ME)
dc.language.iso	en_US	en_US.UTF-8
dc.relation	https://opus.lib.uts.edu.au/bitstream/10453/197221/1/thesis.pdf
dc.rights	info:eu-repo/semantics/openAccess
dc.rights	The author owns the copyright in this thesis including all reproduction and reuse rights for the work. The work may not be altered without the permission of the copyright owner. Attribution is essential when quoting or paraphrasing from this thesis.
dc.rights	© 2025 Zichen Xu
dc.rights	au.edu.uts.lib/cph
dc.title	Nuclear-targeted CRISPR Delivery By Using Aptamer-modified Polymer Lipid Nanoparticles	en_US.UTF-8
dc.type	Thesis
utslib.copyright.status	open_access	*

Abstract:

Efficient clinical translation of CRISPR/Cas9 gene editing requires delivery platforms capable of overcoming intracellular barriers and promoting nuclear localization. In this study, aptamer-functionalized polymer-lipid hybrid nanoparticles (PLHNPs) were developed as a nuclear-targeted non-viral delivery system for CRISPR/Cas9 in cancer cells. The nanoparticles consisted of a biodegradable PLGA core and a lipid shell conjugated with the AS1411 aptamer, which targets nucleolin overexpressed on the membrane and in the nuclei of many cancer cells. This design was intended to enhance both cellular uptake and nuclear transport. The PLHNPs were prepared by nanoprecipitation and optimized for size, polydispersity, surface charge, and loading efficiency. Aptamer conjugation was confirmed by FTIR and zeta potential analysis. In MDA-MB-231 and HCT-116 cells, AS1411-modified PLHNPs showed enhanced uptake and effective nuclear accumulation of CRISPR/Cas9 complexes, as demonstrated by 3D confocal imaging. Functional studies targeting Lcn2 showed increased indel formation, reduced gene expression, and suppressed cell migration. Compared with non-targeted systems, the aptamer-modified PLHNPs achieved approximately 30% higher editing efficiency.

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http://hdl.handle.net/10453/197221