Before joining the CCSF Biotechnology Program, I had completed two Masters degrees and worked as a research assistant working on Lupus. After having taken a long career break to care for my family, I hoped to re-enter the workforce as a research associate in the field of cancer therapeutics. I came across the program while looking for internship opportunities to improve my capabilities with newer biotechnology tools and techniques. The program refreshed my skills, deepened my understanding of molecular biology concepts, and helped me build a network with people in the biotech industry. I plan to utilize the skills I have acquired to effectively position myself in the biotech industry.

I completed my CCSF Bioscience Internship at CODA Biotherapeutics working with AAV (adeno associated viral) vectors to create gene therapies for neurological disorders. My project involved producing high quality recombinant AAV vectors that optimize the capsid selection (tissue specific targeting) and cassette selection (receptor expression) that will be used for in vivo and in vitro studies. To do this I employed mammalian cell culture in multilayer flasks, triple plasmid transfections, Iodixanol gradient purification of rAAV vectors, digital droplet PCR (ddPCR) and silver staining techniques. I performed a series of passages of HEK293 cells from frozen vials to multilayer flasks using aseptic cell culture techniques. I then pre-seed the cells for triple transfection with the plasmid of interest, repcap and helper plasmids into AAV293 cells. After 72 hours, the recombinant AAV vector was harvested using PEG and benzonanze chemicals. The vector harvest was then purified using iodixanol gradient ultracentrifugation and viral preps to concentrate the vector. The purified vector was then tested for concentration by droplet digital PCR and tested for purity by silver stain technique. The purified vector was then aliquoted and stored at -80 degrees for further in vivo studies. 

I hope to be able to use my understanding of techniques in cell biology and molecular biology in AAV vector production to pursue a meaningful and promising research career in gene therapy. I hope my research will help future scientists to produce high quality recombinant AAV vectors that can be used to deliver therapeutic agents in patients with neurological disorders like epilepsy and neuropathic pain.