Ayusman Dash

Ayusman Dash

Dept. of Cancer Biology
Vontz Center for Molecular Studies
3125 Eden Ave
Cincinnati, Ohio 45267
Email dashaa@ucmail.uc.edu

Professional Summary

ORCID ID: 0000-0003-1394-3174

Education

PhD: University of Cincinnati, College of Medicine 2022 (Cancer and Cell Biology Graduate Program)

Research and Practice Interests

Regulate PRPS1 (rate limiting protein for nucleotide biosynthesis) expression with single base mutations using CRISPR-Cas9 to treat developmental and metabolic disorders. Investigate cellular senescence and mitochondrial dysfunction caused by disrupted PRPS1-bound YY1 signaling as a consequence of these mutations.  

Technique/Experimental expertise:

  1. CRISPR-Cas9 (Adenine/Cytosine Base-Editing, ssODN based HDR, Prime Editing, NHEJ) to create base edits/deletions/knockouts in Mouse Embryonic and Human Lung Fibroblasts.
  2. Mammalian cell culture: Fibroblasts (NIH-3T3, TIG-3-20, BJ), Epithelial cells (RPTEC, MCF7), Lymphoblasts/Immune Cells (RS4;11, CCRF-CEM). Cell Counting (Hemocytometer/Countess II FL Cell Counter).
  3. Created workflow for CRISPR gRNA (cloned into pLKO-BfuAI-pGFP) + Cas9 (ABE7.10, ABE8e, NG-ABE8e, VRER-BE3, dCas9, hSpCas9, hCas9_D10A) transfection->fluorescence based single cell sorting->screening for pure clonal mutant expansions.
  4. Fluorescence Assays (Alamar Blue, Hoechst, CyQUANT, CM-H2DCFDA), Luminescence Assays (AMP-Glo Luciferase, Dual-Glo Luciferase), Absorbance Assay (BCA) using CLARIOstar. Kinase Enrichment Assay (ActivX), Proximity Labeling Assay (TurboID), Click Chemistry (Click-IT AHA) in mammalian systems.
  5. Plasmid/gRNA/primer design, cloning and optimization, Restriction Digest, Bacterial Transformation, Golden Gate Assembly.
  6. Transfection/overexpression (Lipid-based/Electroporation), Lenti/Retro-viral infection and screening, shRNA Knockdown.
  7. Flow Cytometry (Fortessa, FACSAria), Brightfield and Fluorescence Microscopy.
  8. Molecular Biology: PCR/RT-qPCR, DNA/RNA extraction, Electrophoresis (Agarose, RNA denaturing, SDS-PAGE, Native PAGE), Western Blotting, Immunoprecipitation.
  9. In vivo: Mice Breeding, Tissue Harvesting, Genotyping and Euthanasia.
  10. Seahorse XFe96 Analyzer, Size Exclusion Chromatography (SEC).
  11. Protein Engineering: modeling and design using PyMOL, experience with recombinant proteins.
  12. Created Lab’s antibody, cell line, protein, RNA, glycerol stock databases.
  13. Sample preparation for FRET (FLIM) and Super Resolution Fluorescence Microscopy (SIM).
Computational skills:
  • PhD: Biostatistics (SigmaPlot, GraphPad Prism)
  • BS-MS Dual degree/High School: Perl, C, Python, JAVA
Software/Browser expertise:
  • Benchling, SnapGene, Adobe Illustrator, FlowJo, Microsoft package, Seahorse Wave, Image Lab, ImageJ, Integrative Genomics Viewer, UCSC Genome Browser, Ribo-seq genome browser, Ensembl genome browser, BioRender, WebLogo.
In Silico Workshops Attended:
  • 2020: BIOF 087 | Programming for Biomedical Researchers workshop organized by Foundation for Advanced Education in the Sciences (FAES) at National Institutes of Health (NIH).
  • 2018: UCSC genome browser workshop, University of Cincinnati | Libraries.

Contact Information

Academic - Dept. of Cancer Biology
Vontz Center for Molecular Studies
Cincinnati  Ohio, 45267
dashaa@ucmail.uc.edu