The CIRM SPARK Lab Experience by Yasmine Mahmoudieh

IMG_4370I’ve always been interested in becoming a doctor and directly helping patients. Until the CIRM SPARK Lab experience I did not realize how important research is to developing treatments. I now have a greater appreciation for the work that researchers do and an understanding for what is required to conduct research.

On the first day, I was assigned to the Precision Neurogenetic group. Working alongside my mentor Kyle Fink and four graduate students Jasmine Carter, Sakereh Carter, Peter Deng, and Julian Halmai has been the most gratifying experience of my life. Their passion for the field is clearly evident and it emanates in all of the work they do daily at the lab. All five of them have played an immense role in making this experience one that I will always remember.

For the past 8 weeks, I have shadowed them and learned about their research to target and modulate transcription of genes associated with neurological diseases by creating artificial transcription factors which are DNA binding domains fused with effectors that target regulatory regions of that gene. They use this technology to work towards the potential treatment of diseases such as Rett Syndrome, Huntington’s disease, juvenile Huntington’s disease, Angelman Syndrome, Glioblastoma, and CDKL5 deficiency. Traditional gene therapy has shown promise in early stage clinical trials to treat several genetic diseases. This is accomplished due to the addition of a known gene that is lacking in these diseases. In recent years, advancements in technology have allowed for genome modification and drastically altered the biomedical field.img_4371.png

My personal project was trying to optimize transfection efficacy to better integrate gene editing technologies for potential treatment of neurological diseases. Transfection is the integration of foreign DNA into cells using a non-viral method. I transfected SHSY5Y neurons and HEK293 kidney cells with a transcriptional activator VP64-dCas9-VP64, containing eGFP in multiple disease cell lines using the three commercial reagents DNA-In, Lipofectamine 3000, and PEI. I varied the amounts of DNA and reagents to determine how I could obtain the highest transfection efficacy. Through my experiments I was able to determine that the PEI reagent resulted in the highest number of GFP positive cells in both cell lines tested and transfection with the PEI reagent can be further optimized by using different concentrations corresponding to DNA concentrations.

During my internship as a CIRM SPARK Lab student, I have transfected two different cell lines, grown a cell culture, used the sonicator to homogenize brain tissue to extract RNA, done a mini prep to extract plasmid DNA, worked on the Nikon TiU inverted fluorescent microscope, used a flow cytometer, isolated DNA, and completed a 14-day sterility assay in the Good Manufacturing Practice (GMP) Facility. The skills I have accumulated at the UC Davis Institute for Regenerative Cures are ones I will be forever grateful for as I pursue a career in the medical field.

 

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