Team:TP-CC San Diego/Results
Results
Effects of ecDNA
There is a significantly depressed rate of cell growth due to the treatment. New cell growth, measured in the increased of density, was significantly retarded in the induced group when compared to the control.
Parts
BBa_K2369000
BBa_K2369001
BBa_K2369002
Images
EGFR copy number
EGFR vs. Normalized Vector Expansion
E1G4
Gels
Digested TLCV2 Vector
PCR
GBM39 Genomic DNA
References
Guide RNA plasmid design
12 Well Plate Design
100 Basepair ladder
STBL3 competent cell colonies containing guide RNAs
E1G2
E1G4
E8G2
E8G3
Summary
We were able to confirm that the GBM39 cell line had a substantial copy number of the EGFR gene. From there, we designed 4 guide RNAs. Two from exon 1 and two from exon 8. We were unable to design any gRNA using exons 2-7 because in the GBM-39 cell line, that portion of the EGFR gene is mutated. Using the TLCV2 backbone, we removed the stuffer sequence and inserted the guideRNAs in its place. We were successful with constructing 3 gRNAs but the 4th failed to clone twice. After Sequencing our plasmids, we transfected the guideRNAs into GBM39 cells. Currently, we are monitoring the growth of the cells, and confluency and growth of the cancerous cells has been, over the past 2 months, negatively affected in a statistically significant manner - with over a reduction in total confluency growth of over 50% when compared to the control, suggesting that our treatment has an inhibitory effect upon cancer cell growth.
