Nicholas Willis Email & Phone Number

Boston, Ma, Us

Somerville, Massachusetts, Us

As RNA Therapeutics Core Lab Manager, I provide both operational and experimental support to a small team bringing best-in-class circular RNA products to academic and biotech communities of greater Boston and beyond.Bench Scientist Duties: support development of new processes for RNA and LNP asset production, circRNA product functional validation and characterization. Molecular and cellular biology support to the RTC team and visiting entrepreneurs in residence.Core Manager Duties: day to day operations control, CapEx equipment purchase and onboarding, consumable supply and stock management, tissue culture surveillance and standard lab practice enforcement, facilitate sample intake and tracking, working group member for GMP facility buildout, GCTI team checkin.

I provide molecular and cellular biology expertise and on-site experimental design and support as needed, augmenting client company in house teams. Activities include new target identification, target validation, platform support, MOA studies initiation, and execution of various endpoint assays. Hardware provided: Lonza nucleofector 4D and X-unit.I have worked with seed and small biotech companies on experimental design to delete improve use of CRISPR technology to delete or target genes of interest or provided direct bench support to study MOA of lead assets using a combination of chemical genetics and molecular biology tools (Nexo Tx and Tasca Tx).

Lexington, Ma, Us

• Managed/trained direct hires (senior scientist and RA) expanding Platform capability and capacity.• Established in house CRISPR mediated gene-editing platform supporting all pre-clinical research.• CRISPR Platform crucial to rapid and robust target validation and de-risking studies, pharmacodynamic biomarker candidate evaluation, and target selectivity assays for CYT-0851 (clinical stage MCT1 inhibitor).• Engaged in CYT-0851 MOA studies using a variety of cell-based assays and internally developed genetic tools.• Established a variety of cell-based assays to measure DNA DSB repair efficiencies, DNA repair protein responses, and physical measurement of DNA damage.• Developed a unique target engagement assay critical to chemistry SAR and screening in support of new target program nomination (MMEJ repair protein Pol Theta, POLQ).• Established in house DNA damage quantification assay (comet assay) to characterize lead chemical matter across the DNA Damage Response (DDR) pipeline. • Managed flow cytometer operation, expansion and training.• Liaised closely with CROs developing molecular biology reagents for cell-based assays. • Liased with CRO initiating siRNA screens to expand potential indications for preclinical programs or to generate reagents important to nascent programs.• Core member of target identification working group; new target ID/evaluation, followed by genetic validation and de-risking studies.• Core member pipeline working group; 20+ IP in-licensing evaluation with data room due diligence.• Summarized data, Platform objectives and technology development routinely to project & management teams.

• Conceptualized projects leading to $2M funding and publications in PLoS Genetics, NSMB, & Nature, STAR Protocols, Current Opinions Genetic Development, Molecular Cell, Methods in Molecular Biology.• Trained incoming staff: two postdoctoral fellows, one assistant in tissue culture.• Identified DNA tandem duplications (TD) as a biomarker of BRCA1 loss in cancer.• Defined a protein network regulating TDs revealing the mechanism of TD formation.• Expanded the Tus/Ter HR reporter system toolset with 24+ reporter variant lines.• Generated Tus/Ter reporter in human immortalized and cancer-derived cell lines.• Completed an initial siRNA-screen exploring the roles of ~210 target genes in stalled fork repair.• Established CRISPR/Cas9 plasmid, in-house sgRNA transcription, and RNP use to delete/point mutate/tag target genes in ES cells.• Utilized CRISPR/Cas9 for use in molecular cloning, and as a nuclease to induce site-specific HR/repair repair in vitro.• Completed a survey of 292 BRCA1 VUS ("variants of uncertain significance") alleles for rescue of BRCA1-mutant cell HR repair defects and PARP inhibitor sensitivity in collaboration with Dr. Jonkers group (NKI).• Provided data critical for ongoing and completed patent applications.

Boston, Ma, Us

• Produced key data for $1.5M grant funding and publications in PNAS, PLoS Genetics, Trends in Cancer, Molecular Cell, Nature Communications, and Nature.• Patent issued: screening of BRCA1 variants of uncertain significance, USPTO #10697026• Optimized and validated cell model systems to measure DNA DSB repair.• Established 2D Southern blotting measuring Tus/Ter-induced replication fork stalling.• Evaluated the roles of Fanconi Anemia gene family proteins, BRCA2, and RAD51 in stalled fork repair.• Provided critical data for RO1 and R21 funded applications focused on BRCA1 VUS alleles' HR repair defects and chemotherapeutic sensitivities.

Boston, Ma, Us

• Pioneered/validated the Tus/Ter stalled fork repair reporter to quantify homologous recombination (HR) induced by site-specific replication fork stalling on the mammalian chromosome.• Revealed a key regulatory role for BRCA1 in stalled fork repair.• Optimized and validated cell model systems to measure DNA DSB repair.• Established 2D Southern blotting to quantify replication intermediates upon Tus/Ter-triggered fork arrest.

Boston, Massachusetts, Us

• Utilized DNA combing to explore checkpoint-dependent regulation of replication origin firing and replication fork rate in response to MMS, hydroxyurea, and bleomycin.• Explored the role of PCNA SUMO and ubiquitin modifications in replication slowing in response to MMS.

Cambridge, Ma, Us

• Developed murine models of human cancers with Dr. David Tuveson in Dr. Tyler Jacks' lab.• Engineered stem cells and mice carrying Cre-LoxP mediated conditional mutant alleles of the tumor suppressors p53 and K-ras.• Measured p53 and K-ras mutant contribution to cancer development using mouse aging studies.• Determined impact of these mutations on cell physiology and behavior in mouse fibroblasts in vitro and in vivo.• Investigated and published on the efficacy of STI571 (Gleevec) treatment on GIST primary cells in vitro.• Published several papers describing mutant mice as models for human pancreatic and lung cancers.

Bar Harbor, Me, Us

• Explored the mechanisms of vertebral column formation during mouse development. • Generated detailed genetic maps for three spontaneous mutations affecting axial patterning and skeletal formation, specifically Danforth's shorttail (Sd), Pintail (Pt), and Truncate (Tc).

Orono, Me, Us

Assisted Dr. Dorothy Croall characterizing the protease calpain and its role in calcium-dependent cellular functions. Synthesized cDNA libraries from total RNA (produced by starving cultures of Dictyostelium discoideum) and screened those libraries for homologues to murine calpain.

Orono, Me, Us

• Constructed a mutant gene library to characterize G-protein signaling pathway(s) essential for the completion of the life cycle of the slime mold Dictyostelium discoideum.• Capstone, senior research project characterizing several mutants of the G protein subunit Ga2 in signaling.

Ph.D., Biomedical Sciences

B.Sc., Biochemistry