Difference between revisions of "Team:TP-CC San Diego/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.