Publication Date
2026
Presentation Length
15 minutes
College
College of Sciences & Mathematics
Department
Biology, Department of
Student Level
Undergraduate
Faculty Mentor
Rebecca Adams
Presentation Type
Talk/Oral
Summary
In eukaryotic cells, the transport of mRNA from the nucleus to the cytoplasm is a very regulated process controlled by nuclear pore complexes (NPCs). During stress conditions, such as heat shock, most mRNA export is inhibited in Saccharomyces cerevisiae, yet the transcript for SSA4, a gene responsible for refolding denatured essential proteins, is exported. The mechanism behind this selective export remains unknown. To investigate this, we used CRISPR-Cas9 to generate a reporter strain in which the SSA4 open reading frame was replaced or inserted with green fluorescent protein (GFP). This reporter is designed to produce GFP only under conditions that promote SSA4 export, such as heat stress, and not in export-deficient mutants (e.g., nup42Δ). The GFP gene was amplified via PCR to give us a product that we inserted into the SSA4 locus using CRISPR-Cas9. Successful integration of GFP genes was confirmed by PCR, and GFP expression will be checked using fluorescence microscopy. This tool will allow SSA4 expression to be visualized, opening the door for future experiments aimed at identifying the SSA4 sequence elements necessary for selective export.
Recommended Citation
Khairollahi, Aara and Adams, Rebecca, "Generating a GFP Reporter via CRISPR to Visualize SSA4 Export" (2026). SPARK Symposium Presentations. 881.
https://repository.belmont.edu/spark_presentations/881
Included in
Cell Biology Commons, Genetics Commons, Molecular Biology Commons
