Date of Graduation


Document Type


Degree Name

Bachelor of Science in Biomedical Engineering

Degree Level



Biomedical Engineering


Muldoon, Timothy


CCL2 is an inflammatory mediator that is released by tumor cells to activate and direct immune cell species, especially macrophages, to inflammatory sites within the body. The goal of this project was to successfully generate knockout the CCL2 ligand gene using a CRISPR/Cas9 complex delivered via lipid mediated transfection. The sgRNA and Cas9 mRNA were introduced into the cells via lipid-mediated transfection. The cells were incubated for 4 days, before being analyzed using PCR and gel electrophoresis. We expected to see one band on the first gel and two bands on the second gel. Two bands appeared on the first gel and 2 bands appeared on the second gel. This indicates that the target sequence was amplified, but the transfection efficiency was too low for the enzyme to detect cleavage. We conclude that lipid mediated transfection is an efficient method for introducing foreign genetic material into CT-26 cells.


CCL2, CRIPSR, Cas9, transfection, sgRNA