Development Training
Resume:
Mailin Van Hoosear
Gainesville, FL 32608
ac5ri2@r.postjobfree.com
EXPERIENCE
AGTC, Alachua, FL Jan 2016-present
Research Associate IV, Preclinical Research and Development Wet age-related macular degeneration, wet AMD.
Dry age-related macular degeneration, dry AMD.
X-linked retinitis pigmentosa, XLRP.
ALD/AMN
Cell based assay development to validate potency of AAV-therapeutics. Familiarity with AAV production by triple transfection and herpes simplex methods and AAV quality control assays. AAV tittering by quantitative PCR.
Writing of technical reports, data generation and preparation of poster presentations and research manuscript. Cloning of gene of interest ABCD1 in pTR-vector backbone. Expression ELISA development, AAV2-ABCD1. Collaboration with outside vendors to characterize protein product of AAV therapeutic. Evaluated stability of cDNA and RNA of AAV therapeutic by sequencing post-transduction genomic DNA and reverse transcribed RNA. Successful amplification of reverse transcribed full-length (difficult) transcript, RPGRORF15. Characterization of RPGR protein product by immunoprecipitation, LC-M/MS(in collaboration with UF Dept of Chemistry), and post-translation modifications.
PCR quantitation of microRNAs across species.
Lab Management duties:
Prepared standard operating procedures (SOP) for laboratory housekeeping, gowning and materials transfer. Prepared standard operating procedure for multimode plate reader. Reviewer of additional assay development or operations SOPs. Training of junior associates, summer students; instrument evaluation, demo and purchasing decisions. Instrument service and repair directly with vendor.
Maintained well-stocked consumables in research and development laboratory. Ordering of research reagents and consumables for the research and development team. Completed technical report writing, SOP writing, quality assurance, quality control, and good laboratory practice training workshops. Autoclaving of biohazardous waste, monitoring laboratory sinks, logbook maintenance, shipping and receiving of reagents and other materials. Novartis Institutes for Biomedical Research, Cambridge, MA Scientist II Neuroscience Jan 2013-Jan2016
Frontal temporal dementia and the role of microglia in neuro-inflammation, functional assay development. Friederich’s Ataxia small molecule drug discovery, functional assay development. Involved in recruiting junior and senior staff, training junior associates, assisted with evaluation and purchase of instruments, organization and relocation of a new research division.
Novartis Institutes for Biomedical Research, Cambridge, MA June 2010-Jan 2013 Scientist I, Developmental and Molecular Pathways Department Small molecule drug discovery for spinal muscle atrophy (SMA). Post life pharmacodynamic (PD) analysis (transcript and protein) of lead molecule efficacy in mouse model of SMA. Extensive quantitative real time PCR custom assay design to validate next generation sequencing (RNAseq) data. Ex-vivo compound treatment studies for transcript and protein PD analyses in healthy and SMA-patient PBMCs. Ex-vivo compound treatment in SMA model spleen PBMCs for transcript/protein PD analysis. Compound effects on cell cycle by propidium iodide staining and flow cytometry analysis. Over-expression of wild-type and mutant GBA in neuroblastoma to evaluate markers of proteosomal and lysosomal degradation to establish a potential link between Gaucher’s and Parkinson’s disease. Mentored junior associates and an internship student on cell and molecular biology techniques and assisted in organizing their project goals. Proteostasis Therapeutics, Inc. Cambridge, MA Sept 2009-June2010 Associate Scientist III, Unfolded Protein Response Pathway Program Unfolded protein response (UPR) pathway assay development. Analyzed Panomics multiplex gene expression data to examine downstream effects of over-expression of UPR pathway genes in established cell lines.
FRET assay to screen compounds that may inhibit IRE1 endonuclease activity. Secondary validation of active compounds in a UPR-pathway mRNA-splicing luciferase reporter assay. Training junior associates in cell and molecular biology techniques, generation of mammalian over-expression stable cell lines. Novartis Institute for Biomedical Research, Cambridge, MA Sept. 2005-Sept 2009 Scientist I, Apoptosis Program, Developmental and Molecular Pathways Cell death pathway expansion project with a focus on identifying novel interactors of BCL2. Established stable cell lines overexpressing Bcl-2 family members in constitutive and TET-inducible systems, and implemented InCell western
(Licor) technology to streamline the screening of positive clones. Novel targets were followed up in siRNA/shRNA to evaluate cell death phenotypes in cancer cell lines, immuno-precipitation, in-vitro kinase studies were employed to validate kinase interactors. Also participated in early drug discovery for p53-modulated cell death pathways. Contributed to recruiting and training of junior associates. Bionaut Pharmaceuticals, Cambridge, MA April 2003-Sept 2005 Senior Research Associate, Oncology Program
Contributed to the development of Bionaut’s sentinel cell-based reporter system, a platform for drug discovery. Generated stable cell lines using retroviral transduction to introduce GFP or Luciferase reporters into genetic sites regulated by various biological stimuli. Employed reporter cell lines responding to ligand or growth factor stimuli in drug screenings. Investigated the potential mechanism of action of active compounds in the reporter cell lines. Pathways specifically studied in my group included TGF-β and TNF-α. Contributed to the hiring and training of new associates. Praecis Pharmaceuticals, Waltham, MA Aug. 2001-Aug. 2003 Research Associate, Biochemistry Department
Optimized growth conditions of bacterial cell cultures for recombinant protein expression. Produced and purified milligram quantities of recombinant protein by affinity chromatography. Assisted in the development of fluorescent polarization assays to evaluate protein-DNA interactions and compound inhibition studies. Characterized proteins through calorimetric quantitation, Coomassie and immunobloting analysis, immunoprecipitation and gel-shift assays. Engineered the cloning of a new DNA insert to correct bacterial host misread of a stop codon in a recombinant gene construct.
RESEARCH TECHNIQUES
Cell Biology: Mammalian assay development (stable, transient expression), post-life (in vivo-study) sample analysis, microscopy assay development, primary murine microglial and cortical neuron isolations and culture, peripheral blood mononuclear cell (PBMCs) isolations from healthy and patient bio-specimen, extensive mammalian cell culture experience (established and primary tissue), pathway biology. Molecular Biology/Biochemistry: gene expression analysis (Taqman), molecular cloning, recombinant gene over- expression (plasmid, pLenti, AAV), gene knockdown (shRNA, siRNA), molecular cloning, immuno-precipitation, immuno-blotting, HTRF and ELISA assays, custom gene expression (Taqman assay) design, biochemical assay development, metabolic assay development.
TECHNOLOGY
Software
Microsoft (Excel, XLFit, Word, PowerPoint), GraphPad Prism(statistics, figures for publication), Spotfire, Photoshop, PrimerExpress 3.0, Sequencher, Vector NTI, AxioVision (Zeiss microscope), ModFit (cell cycle flow cytometry), FACSDiva.
Instrumentation
SpectraMax i3X, Guava InCyte (FACS), Perkin Elmer EnVision, FACS LSRII, InCell 6000, Zeiss fluorescence microscopy, ABI ViiA7 Ruo quantitative PCR system, ViCell, Spectramax Paradigm, automated cell seeding instruments, LiCor Odyssey imaging, IncuCyte, Nexcellom Auto2000, Prescellys, BioRad ChemiDoc MP, Stratagene Bioanalyzer, Zeiss dissection microscope.
EDUCATION
B.S. Biology Major, Chemistry minor (GPA 3.56) University of Massachusetts, 2000 Harvard Extension Biotechnology Master's level courses PUBLICATIONS
SMN2 splice modulators enhance U1–pre-mRNA association and rescue SMA mice. Nature Chemical Biology 11, 511–517 (2015).
PERSONAL
Fluent in English and Spanish.
Volunteer work
Quincy Hospital, Rehabilitation Unit:
Massachusetts Alliance for Portuguese Speakers (English as Second Language instructor) Boston Living Center (Center for people living with AIDS) Interests: Baking, bicycling, literature, history, technology.
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