Jenna Williams

•Small molecule CFTR modulator treatment of primary human bronchial epithelial cells (HBE) and stable cell line cystic fibrosis bronchial epithelial (CFBE) cells to identify effects on mRNA and protein levels of candidategenes. • Created a comprehensive set of primers and developed the protocols to genotype from genomic DNA to determine the specific mutations and polymorphisms in the exons of CFTR. • Performed metabolite ELISA assays to quantify metabolites from blood, cells and media to characterize those potential altered in cystic fibrosis. • Supported successful IND-enabling studies by performing Ussing assays on primary HBE cells to characterize PTI CFTR modulators PTI-801 and PTI-808 functional activities, as well as supporting pre-clinical programs by providing chloride transport activity for small molecule compound CFTR modulators. • Supported discovery programs by profiling high-throughput hits from cell line screens in the direct measurement of CFTR-dependent chloride transport in primary HBE cells using the TECC24 robotic platform.

• Designed and constructed various plasmids for research in the CAR-T program while doing initial testing for expression of the new CARs and proteins. • Maintained mammalian cell culture for viral vector production and characterization. • Produced small-scale lentiviral vectors and transduced human T-cells in small-scale micro culture and larger in vitro scale production. • Performed assays to determine the characteristics of genetically modified cells, analyzed the data and pointed out concerns and presented findings. • Continually updated and created protocols and procedures for the vector biology team to streamline all support aspects for the CAR-T immunotherapy team.

DNA sample prep for sequencing reactions, operation and maintenance of high throughput robotics and sequencing instruments. Data entry in Laboratory Information Management Systems, retrieval, arraying, culturing and distribution of plasmid clones and analysis of sequencing data to validate sample identity. Involves laboratory techniques including high throughput robotics, DNA sequencing, DNA transformations, plasmid cloning and sequencing analysis. Molecular biology software is used (BLAST, Vector NTI, primer design and LIMS). Presents poster at department wide retreat explaining and detailing service platforms. Works with scientists worldwide to find specific genes of interests and plasmids specific to their research.

Formation of Bacillus Subtilis Cells for Sporulation and Expression Project: Result: Devised successful protocol for sporulation of viable Bacillus subtilis cells able to survive in harsh conditions, showing Green Fluorescent Protein (GFP) fluorescence for 6+ months after storage, with intention of using cells as platform for other riboswitches inserted into field device used to detect harmful environmental substances, such as TNT. Actions: Created cells, transforming into plasmid containing GFP fluorescent reporter protein, sporulating cells utilizing protocol, lyophilizing and storing at room temperature. Selection of TNT Riboswitch Project: Result: Successfully identified several TNT aptamers with high activation ratios to become part of TNT Aptamer Library, after verifying and screening aptamers’ ability to bind to target through Systematic Evolution of Ligands by Exponential enrichment (SELEX) method. Actions: Formulated TNT aptamer with TNT riboswitch, reporter fluorescent protein, and TetA fusion protein, via dual genetic selection. Lab Processes & Skills: • Tissue Culture: transfection of stable cell lines, terminal cell lines, quantitation of antibody with octet. • Molecular Biology: mini-/maxi-prep, western blot for protein, BCA Assay for protein, SDS-Page protein purification, expression of green and red fluorescent proteins. • Cloning: PCR, RT-PCR, gel electrophoresis, quick change, transformation into E.coli, Bacillus subtilis, Bacillus megaterium, and Yeast (YPH499) cells, ligation, vector mapping, infusions, gel purification, PCR purification, restriction enzymes/restriction digestion, In-Fusion cloning, site-directed mutagenesis.

Tissue Cell Culture: transfections, created stable and terminal cell lines to quantitate by octet, sustained cell culture line of monkey claudin cells, rat and mouse cells, passage of CHO-K1 cells Molecular Biology: cloning and optimizing antibody expression in different vectors testing for selection (IRES, Tripartite, Blasticidin, Neomycin), trouble shooting for cloning Started set-up for Ribosome Display Library Developed exceptional troubleshooting skill sets – as summer intern reporting to Mentor daily – to resolve large number of laboratory-related cloning and comparable issues: • Improved laboratory productivity by completing quality research projects on or before deadline, cloning and optimizing antibody expression in different vectors for selection (IRES, Tripartite, Blasticidin, Neomycin). • Initiated Ribosome Display Library Program, performing transfections, creating stable and terminal cell lines quantified by octet, and sustaining cell culture line of monkey claudin cells, rat and mouse cells, and passage of CHO-K1 cells.