- Engineered next-generation CRISPR-Cas9 gene editors with enhanced HDR- Demonstrated proof of concept of a multi-kilobase gene editing strategy in mammalian cells- Designed and executed a pipeline to transfect mammalian cells, extract genomic DNA, and generate amplicons for 1000 samples over 3 months for next-generation sequencing- Developed and optimized a protocol to express and purify a Cas9-based fusion protein from E. Coli using a two-step FPLC purification process- Cloned >50 plasmid using In-Fusion and restriction enzyme cloning- Created company-wide organization systems for oligonucleotides, plasmids, and linear DNA and a database for tracking experimental conditions and corresponding physical samples- Trained and led a team of three research associatesSkills involved:- Molecular cloning: In-fusion, restriction enzyme- Mammalian cell culture (human liver stem cells (hLSCs), adipose-derived stem cells (AdSCs), fibroblasts, HEK293T, H1299, U2OS, K562, Hepa1-6 murine hepatoma)- Mammalian cell transfection- Fluorescent plate imaging (BioTek Cytation 5)- Immunostaining- Bacterial protein expression and purification with fast protein liquid chromatography (FPLC)- single-stranded DNA (ssDNA) synthesis

Worked on establishing a disease model of PACS1 syndrome in C. elegans

Projects:- Characterized the roles of membrane contact site protein Junctophilin in muscles and neurons in C. elegans, discovering an antagonistic interaction between two membrane contact site proteins in neurotransmission (Piggott et al, Genetics, 2020)- Studied the roles of endoplasmic reticulum – plasma membrane contact site proteins in axon regeneration, particularly regenerative axon fusion (Kim et al, eLife, 2018)Other duties:- Mentored and supervised two PhD rotation students and one undergraduate researcher- Presented research data in group meetings at three-month intervalsSkills involved:- CRISPR/Cas9-mediated gene editing- Molecular cloning: Gateway and Gibson assembly- Fluorescence microscopy- Confocal microscopy of live samples, both spinning disk and laser scanning- PCR & gel electrophoresis- RNA purification- qRT-PCR- DNA purification and microinjection- Laser microsurgery- Data analysis- Manuscript preparation- Public speaking and data presentation

Led discussion sections and lab courses for a total of 100 students over three academic quarters.Courses:- Biochemical Techniques Lab- Human Physiology- Molecular Biology of Human Disease

- Purified and crystallized multi-protein complexes for X-ray crystallography, including a BRCC36 - KIAA0157 dimer whose structure was determined to 2.55 Å resolution (Zeqiraj et al, Molecular Cell, 2015)Skills involved:- Protein purification using affinity and size exclusion chromatography- Protein crystallization manually and using liquid-handling robotics (Gryphon) and optimization of crystallization conditions

- Maintained lab supplies and stock solutions and assisted lab members with research tasksSkills involved:- Preparing stock solutions- DNA purification

- Generated stables lines of Syntaxin-3 and Syntaxin-11 shRNA knockdowns in RBL-2H3 cells to study mast cell exocytosis (Bin et al, PNAS, 2013)Skills involved:- Molecular cloning- PCR & gel electrophoresis- DNA purification- Mammalian cell culture- Cell transfection- Western blotting