The Advanced Therapeutics Laboratory

Department of Biomedical Engineering, Vanderbilt University

Principal Investigator: Craig Duvall, Ph.D. | Mentor: Brock Fletcher (NSF Fellow)

SPRING 2019 – PRESENT: gene editing reporter cell lines

CRISPR/Cas9: The gene-editing protein

CRISPR/Cas9 is a very special protein discovered in bacteria that I believe is the future of medecine, therapeutics, and healthcare. 

Coming to Vanderbilt, I was drawn to the prospects of genetic engineering research and its application as research tools and potential solutions to diseases thought to be incurable.

Since joining the Duvall Lab, I’ve had the opportunity to utilize CRISPR/Cas9 in my research investigations and appreciate its power as a molecular tool in the laboratory.

Generating reporter cell lines

Reporter cell lines serve as research tools to understand what’s going in cells following the introduction of a new input, such as a experimental therapeutic platform.

My Summer 2019 project involved using CRISPR/Cas9 to create two fluorescent reporter cell lines that can be used to screen protein-delivering nanoparticles that my mentor currently is engineering.

Laboratory skills developed include bacterial cell culture, plasmid extraction, in vitro transfection, high-throughput screening, and data analysis.

A look at the cells

The cells you are seeing are ones that have been successfully edited with the plasmid DNA (mTmG) expressing red & green fluorescent proteins.

Why are some green and some red? These cells started off not expressing any fluorescence, but once I introduced the plasmid DNA with the red and green protein sequence, the cells that have been successfully edited first produce the red fluorescent protein.

However, I can now introduce a protein like CRISPR/Cas9 that will go in and “cut out” the red protein sequence in the newly-integrated DNA sequence. The cells that have been successfully gene-edited will then become green.

Current work

Narrowing my research interests towards developing reporter cell lines via gene-editing methodologies, I am currently designing new reporter cell lines that will support efficacy testing of the various therapeutic platforms created in my lab.

Fall 2019 – Present: DNA-delivering nanoparticles

Porous Silicon Nanoparticles

Porous silicon nanoparticles (pSiNPs) can be compared to tiny capsules that have the potential to deliver biological-based therapeutics (also known as “biologics”), such as nucleic acids and proteins, around the body.

Currently, my graduate student mentor is working on demonstrating the capabilities of pSiNPs in delivering biologics. To support these efforts, I wanted to look into the possibility of delivering DNA into cells with the pSiNPs. Using polymers developed in the laboratory, I designed a pSiNP platform in attempts to create this nanoparticle.