Cellular Toolkit

Genome Engineering


Challenge
Develop a set of tools to experiment and learn about HPV intracellular trafficking.  

Outcome
I genetically engineered cells using CRISPR/Cas9 technology. With these cells, we characterized two proteins involved in HPV’s ascent from the cell surface to the nucleus.

Impact
In addition to HPV infection, the proteins we studied are involved in processes like cell growth, cancer metastasis, wound healing, and mood regulation. These cells are freely available to the entire scientific research community.

Role
I developed the strategy, engineered, and characterized the cells. After establishing these methods, I mentored an undergraduate student in the creation of her own set of cells.



Gene edited cells. Cell characteristics were observed under a microscope, daily. Here, the middle cell is preparing to divide - the DNA has been replicated and each DNA copy is being pulled to the opposite side of the cell (seen as two dark parallel lines).



Research approach
Before CRISPR/Cas9, gene knock-out was messy, incomplete, and caused many off-target effects. This relatively new gene editing technology allows scientists to more precisely alter genomes and effect proteins in a cell. Cas9 is a protein that acts like a pair of DNA scissors. DNA is a code, composed of A’s, C’s, T’s, and G’s. By programming a complementary code into the Cas9 protein, we can direct the scissors to a precise location in the genome. I used this method to target and remove specific cellular proteins and grow clonal populations of these cells. My methods are published in Scientific Reports (2018) .

Whiteboard sketches on protocol design. I find using a white board helps to explain complex topics. In addition to visual aids, the time it takes to sketch imposes limitations on speedy explanations and gives listeners time to digest and synthesize information.