My name is Brandon Carris and I am a rising senior attending Crescenta Valley high school. I applied for the CIRM SPARK program to better my knowledge in genetic editing and get a scope of the research side of the medical field. My main goal for this program is to get a concrete idea of how research works and how to conduct gene editing.
Dr. Katie Grausam has been assigned as my mentor in the research lab and she has guided me through the basics of gene editing and tissue staining. Dr. Grausam is a postdoctoral researcher at Cedars-Sinai and has immense experience in the research field.
So far I have learned how to conduct immunohistochemistry in mouse brain tissue with special antibodies that link to genes that Dr. Grausam has edited previously. She knocked out the three genes Pten, Nf1, Trp53 which inhibit tumor growth, thus by knocking them out, tumor could now grow with increased ease. The reporter protein, green fluorescent protein (GFP), was also added to signal where the genes have been edited when analyzed under a microscope. We took two coronal slices of the brain and bathed them in PBS and multiple times. A primary and secondary solution of GFP, Cy5, Iba1 antibodies and PBST were mixed and applied to the tissue to rest. After the 2-day process, the tissue was positioned onto slides and analyzed under a microscope. We analyzed the choroid plexus area where anti-inflammatory genes have been knocked out of the DNA sequences. Three colors were displayed on the microscope: green, blue, and red. The saturated green sections are cells that certainly have had their genes knocked out and replaced with a new plasmid that has the GFP which is illuminated by the microscope. The blue colored cells appear on the microscope because of the antibodies Iba1 and Cy5 which both hook on to microglia and other miscellaneous macrophages. The red colored cells are the mTomato genes that are expressed because of the gene Rosa26 mtmG in the test subject mice that were used. Multiple precautions were exercised to reduce the possibility of contamination of our experiment.
Looking back at my first week at Cedars-Sinai, my experience has been overwhelmingly positive. Dr. Grausam has been extremely helpful and resourceful, always answering my questions, and teaching me the art of research. Whenever I meet a new employee, intern, or post doc, they are always welcoming and happy which makes me love this place even more. Ever since the beginning of high school I always asked myself about what I was going to do in the future. Now that I have spent a week in the program and the lab, I think I might have just found the answer to my question.
RMI CIRM SPARK 