Start Date
2021
Description
To better characterize the fluorescent detection systems, we compared protein localization with ER structure and dynamics for two different methods: (1) transient overexpression, and stable overexpression. The two resident proteins used were Calnexin and Calreticulin, which were varyingly expressed in monkey kidney Cos7 cells and cervical HeLa cells. We begun work on generating a system that would allow for endogenous protein tagging via CRISPR/Cas9 gene editing system. This is a system for knock-in of fluorescent protein coding sequences to endogenous genes of interest within the cellular genome itself. Successful knock-in of the fluorescent protein will allow us to visualize ER dynamics and structure without the added experimental requirements inherent for lipid-based transfections.
Recommended Citation
Adjei, Eyram; Westrate, Laura M.; and Arnoys, Eric, "Visualizing the ER without stressing it out" (2021). Summer Research. 12.
https://digitalcommons.calvin.edu/summer_research/2021/Posters/12
Included in
Visualizing the ER without stressing it out
To better characterize the fluorescent detection systems, we compared protein localization with ER structure and dynamics for two different methods: (1) transient overexpression, and stable overexpression. The two resident proteins used were Calnexin and Calreticulin, which were varyingly expressed in monkey kidney Cos7 cells and cervical HeLa cells. We begun work on generating a system that would allow for endogenous protein tagging via CRISPR/Cas9 gene editing system. This is a system for knock-in of fluorescent protein coding sequences to endogenous genes of interest within the cellular genome itself. Successful knock-in of the fluorescent protein will allow us to visualize ER dynamics and structure without the added experimental requirements inherent for lipid-based transfections.