Abstract

Fundamentally altering cellular function at a genetic level is a major area of interest in the biologic sciences and the medical community. By engineering transfectable constructs that can be inserted to dysfunctional cellular systems, scientists can mitigate aberrant genetic behavior to produce proper molecular function. While viral vectors have been a mainstay in the past, there are many limitations, particularly related to safety, that have changed the focus of genome editing to incorporate alternative methods for gene delivery. Lance Array Nanoinjection (LAN), a second-generation microfabricated transfection biotechnology, is one of these alternative technologies. LAN works by utilizing both simultaneous electrostatic interaction with molecular loads and physical lancing of hundreds of thousands of target cell membranes. The purpose of this work is to demonstrate LAN in the context of in vitro transfection of immortalized culture cells and primary cells. As part of that exploration, three distinct areas of investigation are considered, which include: characterizing environmental factors that impact LAN transfection, demonstrating LAN genetic modification of immortalized HeLa 229 culture cells using an indicator marker, and lastly, investigating the effects of LAN on human primary, neonatal fibroblasts.

Degree

PhD

College and Department

Ira A. Fulton College of Engineering and Technology; Mechanical Engineering

Rights

http://lib.byu.edu/about/copyright/

Date Submitted

2016-03-01

Document Type

Dissertation

Handle

http://hdl.lib.byu.edu/1877/etd8346

Keywords

Lance Array Nanoinjection, saline solution, lance geometry, carbon coating, transient low temperature, injection speed, serial injection, propidium iodide, CRISPR-Cas9, primary fibroblast, PDGFR-b, wound healing

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