Lucinda Tam

• Manage outsourcing of custom genetically modified murine model projects with multiple CRO’s to provide mouse transgenic services in a full-scale (from design, guide and monitor timelines, review QC testing to the shipment and delivery of murine models) and fast-paced environment to support Genentech’s basic and preclinical research programs • Serve in a cross-functional and highly collaborative role and interact both inside as well as outside of Molecular Biology. These interactions include Animal Resources, Research Contracts, Finance, Procurement, production supervisors, animal imports team, Scientists, CROs/vendors, etc in order to coordinate and advance the projects. • Assist in the planning and execution of projects with external vendors while adhering to budget, scope, and schedule requirements. Activities would include preparation summaries, communication, protocols, and quantitative analyses. • Successfully designed more than 200 projects for research models and generated targeted ES cell clones for more than 100 projects resulting in successfully delivering genetically modified murine models for Scientists to study the effects of the desired gene modifications. • 15+ years of gene targeting design experience, Embryonic Stem cell culture and genotyping experience and have transitioned our current lab to the use of CRISPR/Cas9 technology in parallel with Embryonic Stem cell approaches resulting in a quicker timeline to deliver the models to Scientists to start their study and experiments.

• Strong organizational skills, including the ability to monitor and track multiple projects with global and domestic CRO partners resulting in effectively managing the large volume of projects in the pipeline. • Leverage CRO capabilities to generate and deliver custom murine models to Scientists to study the effects of their desired gene modifications. • Meet regularly with stakeholders to assess the state of ongoing projects, identify current or projected problems, proactively suggest solutions, and align with internal and external stakeholders while managing project scope and deliverables. • Work with Research Contracts, Procurement, and CRO in establishing SOW, workflow, design, and review CRO project design, help CRO screen for off-targets in CRISPR projects • Designing and ordering of often very complicated gene targeting vectors (mainly using CRISPR technology if possible) for the manipulation of the mouse genome in Embryonic Stem (ES) cells, to support all of Research’s needs for genetically engineered murine models to facilitate drug discovery • Hired, managed, and trained contractors in cell culture and genotyping

Construct Design (CKO, CKI, KI) using various genome browser websites: Ensembl, UCSC, Vista for ES cell approach • CRISPR gRNA design for simple point mutation KI and KO using Benchling • Mouse models via Embryonic Stem Cell technology • Cre/flp system in ES cell using RGD-Adeno Virus transduction • ES cell derivation/generation from existing mouse colonies • PCR strategies for long/short range, and nested PCR to detect homologous recombination. • Use of RT-PCR Taqman assay to detect neo copy number in targeted ES cell clones • Use of Digital Droplet PCR technology to detect chromosomal abnormalities ( Y chromosome) • Restriction enzyme digest of targeting construct plasmid DNA and preparation of DNA for electroporation • Genotyping of germline mice tail DNA, sequence confirmation and reporting of results in CMS Data base • Primer design using the following programs: GseqEdit, Oligo5, Primer Express, Ensembl and vector NTI vector • CHEF-DRIII Pulsed Field Electrophoresis System for preparation and submission of BAC, piggybac DNA for microinjection • Mini/Midi/Maxi Prep plasmid DNA via Qiagen kits. • Transformation of DNA plasmid with JM109 competent E.coli cells • TA topo cloning • Quantification of DNA using nanodrop • 3730 DNA analyzer for high throughput genetic analysis

Maintained and passaged 6 different human cell lines in T-175 flasks and rollerbottles. Computer analysis of microscope images using Image Software. Count cells for passaging and platting using Guava Via count, from CytoSoft software. Plated cells to 384 well imaging plates using Multidrop 384 machine. Maintained a library of cell lines by freezing them for long term storage in cryopreservation. Used MS Excel to calculate the amount of cells passaged, the amount of cells platted and the amount of cells frozen in each vial for long term storage. Used MS Excel to keep records of stocks received, aliquoted, working solutions, and the cells that are thawed. General maintenance of tissue culture laboratory.

Screening of site-directed mutations in NFkB binding sites in Human beta defensin 2 (HBD2) promoters. Comparison of CCL20, HBD2 gene expression in different treatments of human and mouse airway epithelial cells.

Completed summer project on testing the upregulation of tissue factor protein expression in THP-1 and HuVEC cell lines with different doses and time intervals of TNF-alpha and lipopolysaccharide (LPS).

Tested the anti-microbial effect of HBD6 in airway epithelial cells. Induced overexpression of HBD6 peptide in yeast.