Alexander Cruz

• Independent experimental design and execution to characterize CNS disease targets and their effects; including investigation of functionality, efficiency, impact on gene expression, downstream effects, model relevance, and systemic cell impact
• Developed multiple stable cell lines through lentiviral transduction to investigate which cross-functional diseases can cause similar gene and protein defects
• Clonal selection and maintenance to isolate desired cell populations
• Screened oligos in disease models to determine editing efficiency with NGS sequencing as well as functional disease recovery
• Management and ownership of multiple simultaneous projects for proof of concept, understanding biological feasibility, and recommendations towards program advancement

• Assessed immortalized and primary CNS cell lines with multiple transfection conditions; including lipid reagents, nucleofection, and electroporation for optimized screening of oligos
• Developed protocols based on literature to test novel in vitro disease models and understand their effects on target splicing and protein function
• Characterized cell line disease relevance by investigating protein aggregation and recovery through drug spike-ins and mutated targets
• Utilized Jess Simple Western for analysis and quantification of soluble and insoluble proteins, as well as mis-localization in nuclear vs cytoplasmic fractions
• Lentiviral transduction and plasmid transfection of potential advantageous targets to generate stable cell lines to be used in routine screens and functional assays.

• Responsible for routine screening as well as assay development to grow our platform and screening capabilities
• Assisted in advancement of High Throughput Screening (HTS) protocols through deployment of new automation equipment and novel process development from 96-well to a 384-well plate format
• Created a standard protocol to onboard new cell models; quantifying target expression levels, high-throughput compatibility, transfection efficiency, and reproducibility
• Onboarded new immortalized, primary, and iPSC derived cell lines to adapt into our HTS platform
• Collaborated with chemistry and data science teams for oligo optimization, and platform biology for cell model optimization and troubleshooting
• Assessed transfection efficiency on new cell models, including CNS cell lines, through sequencing, qPCR, and Flow Cytometry
• Trained all new HTS hires in cell culture, transfection, automation techniques, and data analysis

Routine Screeening Responsibilities:

• Transfected target and enzyme plasmids into cells to create disease models
• Transfected multiple large batches of oligonucleotides into immortalized cells for first pass screens, and primary cells for secondary screens
• Conducted RNA isolation through RT-PCR to generate target product for Sanger Sequencing
• Utilized sanger sequencing to conduct SAR analysis and regularly present findings across all company departments

Additional Responsibilities:

• Upkeep of lab supply inventory and ordering of essential materials
• Conducted a cost analysis of reagents from multiple vendors and determined workflow bottlenecks to choose most efficient processes for screening
• Optimization of transfection conditions, timepoints, and dosing