Research in Dr. Stoppel's lab in the Department of Chemical Engineering at the University of Florida
Evaluating Gene Integration Efficiency in IAL-PiD2, a Plodia interpunctella Cell Line
I have been affiliated with this project since May 2024. The focus of this project is to investigate the efficacy of a PiD2 cell line via the use of CRISPR and transfection reagents.
Insect cell lines have a wide array of potential in the recombinant protein production that can be utilized in therapeutics and vaccine technology. They do not require carbon dioxide or non-room temperature environments to be maintained, unlike mammalian cells. They are also cheaper than mammalian cells. Insect cell lines can also replicate human glycosylation patterns through the use of genetic engineering, which is critical for the proper protein function in human therapeutics with glycoproteins. This ability has not been seen in bacterial or yeast cell lines. Despite these benefits, there are not many commercially available insect cell lines, thus making the research of other insect cell types that can be utilized in cell lines pertinent. IAL-PiD2 cells derived from the wing imaginal disks of Plodia interpunctella, could be a great candidate for an insect cell line and provide a valuable platform for recombinant technology. CRISPR and transfection reagents were utilized to deliver the DNA template to modify the phenotype of the cells to show green phenotype via the production of the mNeonGreen protein and assess the cell line's overall potential in the sustained production of proteins.
I assisted in transforming cells with a CRISPR/Cas 9 complex and assessing the performance of three different transfection reagents from Mirus Bio®(TransIT®-X2, TransIT®-2020 transfection reagent, and TransIT®-Insect). My PhD mentor and I constructed plasmids to produce DNA template for the transfection. I assessed the efficacy of the cell line through fluorescence microscopy, and I analyzed images with a CellProfiler® pipeline to quantify the transfection efficiency of each reagent. I am currently working on comparing these results to flow cytometry data.
