The design, characterization, and application of CRISPR-based tool in human cells

Many human diseases are associated with dysregulated gene expression. A subset of these diseases arise from deviated functional protein expression from homeostatic regimes. Either not enough, or too much, functional protein expression can cause toxic accumulation of metabolites, dysregulate regulatory networks, or myriad other systems necessary to maintain homeostasis. The field of epigenetics was born with the hypothesis that some form of regulation above the genetic level could explain the diverse phenotypic traits observed within a multicellular organism possessing a single genotype. Now we understand that homeostasis arises when the rich interplay of epigenetic mechanisms functions correctly. With our ever growing set of CRISPR-based tools, we are on the precipice of targeted correction of aberrant gene expression. Although many of these tools have been designed, there are still limitations regarding the effectiveness of many gold standard tools with therapeutically relevant human cell types. Work in this thesis was completed to: (1) explore the similarities between natural and synthetic epigenetic engineering as well as understanding the importance and complexities that dictate the transcriptional responses associated with dCas9-based histone acetylation. (2) construct a modular user-defined platform for the precise recruitment of proteins with diverse applications in targeted transcriptional activation, mammalian metabolic engineering, and identifying synergistic interactions of endogenous proteins in situ.

Name thesis:degree_name

Doctor of Philosophy

Level thesis:degree_level

Doctoral

Discipline thesis:degree_discipline

Engineering

Grantor

Rice University

Year dc:date.issued

2023

Author dc:creator

• Cabrera, Alan Cavido

Advisor dc:contributor.advisor

• Hilton, Isaac

• epigenome editing
• CRISPR
• dCas9
• Prediction

Statement dc:rights

• Copyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.

Language dc:language.iso

eng

Handle dc:identifier.uri

https://hdl.handle.net/1911/115261

OAI identifier oai:identifier

oai:repository.rice.edu:1911/115261