BMB-5 Investigating the effects of miR-718 on GOS28/GS28 expression and vesicle movement in SH-SY5Y cells
Abstract
In this study, we focused on miR-718 which serves a critical role in serval different cancers according to published works. MiR-718 has a predicted target called Golgi SNAP Receptor Complex Member 1 (GOSR1), whose protein product, GS28, is a SNARE protein (N-ethylmaleimide-sensitive factor attachment protein receptor) involved in vesicle transport from the rough endoplasmic reticulum (RER) to the Golgi apparatus. We hypothesized that upregulating miR-718 would decrease expression of GOSR1/GS28, which would then lead to reduced vesicle movement. We are investigating the effects of miR-718 upregulation on GOSR1/GS28 expression and the effects of miR-718 upregulation on vesicle dynamics in SH-SY5Y cells, a human neuronal model. We are currently validating that GOSR1 is being targeted by miR-718 using reverse transcription and quantitative PCR (RT-qPCR). We used immunocytochemistry to stain cells with an antibody for GS28 protein to determine GS28’s protein levels in cells, and we stained cells with lysotracker red to analyze vesicle movement using fluorescence microscopy. This project is a work in progress that will demonstrate the importance of miR-718 in regulating vesicle movement in neurons, and hence neural transmission.
Keywords
microRNA, vesicle trafficking, neurotransmission, GS28, GOSR1
BMB-5 Investigating the effects of miR-718 on GOS28/GS28 expression and vesicle movement in SH-SY5Y cells
University Readiness Center Greatroom
In this study, we focused on miR-718 which serves a critical role in serval different cancers according to published works. MiR-718 has a predicted target called Golgi SNAP Receptor Complex Member 1 (GOSR1), whose protein product, GS28, is a SNARE protein (N-ethylmaleimide-sensitive factor attachment protein receptor) involved in vesicle transport from the rough endoplasmic reticulum (RER) to the Golgi apparatus. We hypothesized that upregulating miR-718 would decrease expression of GOSR1/GS28, which would then lead to reduced vesicle movement. We are investigating the effects of miR-718 upregulation on GOSR1/GS28 expression and the effects of miR-718 upregulation on vesicle dynamics in SH-SY5Y cells, a human neuronal model. We are currently validating that GOSR1 is being targeted by miR-718 using reverse transcription and quantitative PCR (RT-qPCR). We used immunocytochemistry to stain cells with an antibody for GS28 protein to determine GS28’s protein levels in cells, and we stained cells with lysotracker red to analyze vesicle movement using fluorescence microscopy. This project is a work in progress that will demonstrate the importance of miR-718 in regulating vesicle movement in neurons, and hence neural transmission.