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Author ORCID Identifier

https://orcid.org/0000-0001-5571-2820

Document Type

Campus-Only Access for Five (5) Years

Degree Name

Doctor of Philosophy (PhD)

Degree Program

Chemistry

Year Degree Awarded

2020

Month Degree Awarded

May

First Advisor

Sankaran Thayumanavan

Subject Categories

Materials Chemistry | Polymer Chemistry

Abstract

CRISPR-Cas9 has emerged as a revolutionary genome editing tool which has the potential to permanently correct diseases caused by malfunctions in DNA. This technique can start a new era of ‘genomic surgery’ customizable to the needs of an individual patient. The biggest hurdle in the clinical implementation of CRISPR is the absence of a biocompatible scaffold that can shuttle this protein-based editing technology to the nuclei of targeted cells without permanent insertion of CRISPR-Cas9 associated gene as seen in the case of viral vectors. Permanent expression of CRISPR components, Cas9 and the single guide RNA can induce unwanted mutations in the cells along with the desired edits. Synthetic delivery platforms can transiently deliver Cas9/sgRNA complex to the target cells in their expressed form to favor on-target DNA cleavage with minimal off-target mutations. The focus of this thesis is to develop different synthetic vectors to complex electrostatically or covalently with CRISPR/sgRNA ribonucleoprotein (RNP) complex so that they can efficiently deliver RNPs to the cellular nuclei and induce optimal genome editing. We have described three different strategies based on mesoporous silica nanoparticles, a charge-neutral reactive polymer and a cationic block co-polymer to efficiently encapsulate CRISPR RNPs and deliver it to the target cells for gene editing application. Our described scaffolds can bind and deliver CRISPR RNPs without the need of any permanent modification on Cas9 and deliver it in a traceless fashion which sets us apart from the approaches typically described in literature.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Available for download on Saturday, May 08, 2021

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