23+ Years of Scientific Training

In vitro cellular systems

• Experience using iPSC-derived microglia for target validation and compound testing.

• Adept at using primary cells for dose-response curve analysis of clinical and experimental compounds.

• Adept at modeling CNS diseases using in vitro systems.

• Isolation and assay development using primary immune cells, including human B cells, monocytes, macrophages, dendritic cells, and T-cells from donor blood. Expertise in transfection, activation, and expansion of B and T cells.

• Generation of knock-out and HDR knock-in cell lines using CRISPR-mediated gene editing

• Expertise in generation of primary mouse neuron (cortical and dorsal root ganglion), neural stem cell, astrocyte, oligodendrocyte, oligosphere, microglia, bone marrow derived macrophage, and fibroblast cultures.

• Isolation and culture of primary human fetal microglia.

• Preparation and compound testing using embryonic and postnatal organotypic mouse brain slice cultures.

• Expertise in using mixed co-cultures of primary neuronal and oligodendrocytes for myelination assays

• Culture, maintenance, and experimental design using several different cell lines.

  • HeLa, COS-7, PC12, B104, 293T, Oli-neu, PC12, HT-29, L929, BMDM, Jurkat, U937, BCLC, Daudi, HepG2, RL, MOLM-13, SK-OV-3, Capan1, NALM6.

In vivo animal models

• In depth analysis of several neurodegenerative mouse models including SOD1, 5xFAD, and twitcher.

• Design and analysis of rodent oncology models using CAR-T therapeutics.

• Nervous tissue dissections, including forebrain, hippocampus, cortical hemispheres, cerebellum, hindbrain, spinal cord, optic nerves, and sciatic nerves.

• Recovery surgeries including stereotaxic injection into CNS and PNS regions.

• Generation and characterization of neuron specific Cre-recombinase transgenic mice.

• Development of floxed mice and use of Cre-transgenics to produce conditional knock-out mice.

• Performed cardiac perfusions and tissue preparation for immunohistochemistry and electron microscopy

• Working knowledge of embryonic and postnatal neurodevelopment, mouse husbandry and colony maintenance, and conventional PCR and Taqman SNP genotyping.

Biochemical assays

• Cell viability assays using PI, CellTiterGlo2 and CytoTox-ONE (LDH-based)

• Transcriptional expression assays using firefly and Renilla luciferase and ß-gal.

Gene expression assays

• Bio-Rad droplet ddPCR system

• PacBio sequencing analysis

• NGS analysis of whole genome libraries

• Whole genome Affymetrix and Agilent microarray systems

• Qiagen RT-PCR profiler arrays

• Taqman based qPCR expression

• Conventional, qPCR and RT-qPCR primer and probe design.

Automation

• Routinely use High throughput solutions including the Hamilton Nimbus workstation and Bio-tek plate washer and pump

CRISPR/CAS9

• Identify CRISPR targets

• Design and test sgRNA

• NGS analysis of CRISPR-cut sites

• HDR directed mutation or knock-in incorporation

• Knock-out or SNP cell line development

Protein Expression assays

• Western Blot using chemiluminescent or fluorescent detection

• iBind system

• Plate-based Immunoassays including ELISA and MSD (single-plex and multi-plex)

• Full Moon Biosystems Protein Profiler arrays

• Working familiarity with Quanterix and Luminex systems

FACS analysis

• Multi-channel sorting of cellular populations using iQue and BD FACSCantoII and analysis using FACSDiva and Forecyte software

Molecular Biology assays

• DNA Cloning and plasmid construction

• Chromatin Immunoprecipitation

• Gel electrophoresis mobility shift assay

• DNA and RNA gel electrophoresis

• Northern and Southern blot

• Enhanced chemiluminescence and LiCor detection of immunoblots

• Protein co-immunoprecipitation

• Protein purification from bacterial cultures

• RNAi-mediated gene knock-down

• 5’-rapid amplification of cDNA ends

• Site directed mutagenesis

Virology

• Adenovirus transductions

• Vector design and capsid engineering for adenovirus

• Retrovirus design and production

• Lentiviral and AAV design

• in vivo CNS and PNS administration and expression.

Immunochemistry

• Cryostat and vibratome sectioning of CNS and PNS tissue

• Histochemical and fluorescent in situ hybridization

• Immunohistochemistry and immunofluorescence of cells or tissue sections.

Microscopy

• High throughput Microscopy using Opera Phenix

• Evos

• Confocal

• Fluorescent

• Phase-contrast

• DIC

• Live- and two-photon microscopy

Computational

• Tableau

• Veeva

• SciNote

• ImageJ

• Microsoft Suite (Excel, Word, Powerpoint)

• GraphPad Prism

• BLAST, ClustalW

• Adobe Suite (Illustrator, Photoshop, Reader)

• Dotmatics

• Basic R and Python programming

• Geneious

• Genevestigator