Results
Through a series of transfection optimization experiments (See Experiments) in three mammalian cell lines, we determined optimized seeding densities, DNA concentrations, and transfection methods. We characterized six sgRNAs targeting three endogenous mammalian genes via qPCR. We improved characterization data for NEU 2016’s CRY2-VP64 part as well as demonstrated that our CRY2-VP64 fusion protein with an additional linker sequence has greater fold induction. We demonstrated the light intensity dependency of the LACE system by measuring fold increase of IL1RN RNA at 1000GS, 1500GS, 1750GS, 2000GS, and 4000GS. We also monitored fold increase of IL1RN in untransfected cells illuminated at various intensities and observed possible light-induction of IL1RN. More data is needed to determine why is phenomenon occurs. To learn more about our experiments and results, see Experiments page.
