In vitro cell based assays, small molecule testing, western blotting

SUMMARY

Cell and molecular biologist with expertise in mammalian cell culture, design and implementation of in vitro and in vivo cell based assays, testing small molecules and viral vectors for targeted therapies, expression analysis study, histology, immunochemistry, advanced microscopy, statistical analysis and general molecular biology skills. Strengths include successful project planning and execution, attention to detail, excellent communication and skills to work in multidisciplinary environment.

RESEARCH EXPERIENCE

2011- Current Children’s Hospital Boston, Boston, MA.

Lead, Research Technologist- Hematology/ Oncology

• In vitro testing of small molecules on leukemia models.

• High throughput cell based assays for screening compounds.

• Protein and gene expression analyses to study effects on epigenetic markers.

2008- 2010 St. Jude Children’s Research Hospital, Memphis, TN.

Senior Research Technologist- Experimental Hematology

• Evaluate various lentivirus constructs to identify safe, effective and long-term gene therapy of the blood disorder, Wiskott-Aldrich syndrome (WAS).

• In vitro assays to study protein expression in human cell lines, patient cells and WAS knockout mouse model.

• In vivo assessment of therapeutic potential.

2005- 2007 University of Tennessee Health Science Center, Memphis, TN.

Post-doctoral Scientist- Anatomy & Neurobiology

• Spatial and temporal expression of genes in cerebellum development- comparing various strains of mice.

• Develop cell based assays to quantify developmental processes.

• Combine immuno-histochemistry and high throughput micro-array analyses for better understanding of gene expression and function.

2001- 2005 Columbia University, New York, NY.

Post-doctoral Scientist- Neuropathology

• Study mouse model for neuronal disorder viz. Zellweger Syndrome.

• In vitro and in vivo assays identify cellular mechanisms leading to defects.

• Extensive primary cell culture, immuno-histochemistry, advance microscopy (confocal, DIC and time-lapse video microscopy).

• Important role in setting up lab, establishing techniques, connections and trouble-shootings.

• Mentored summer students, successfully integrated and supervised their progress in sub-projects.

SKILLS

Mammalian cell culture: Primary cell, cell-lines and organotypic cultures, suspension and adhesive cell cultures, serum free defined medium, isolation and culture of human peripheral blood mononuclear cells, isolation and culture of lineage negative lymphocytes from mouse spleen and lymph nodes.

Cell based assays and Pharmacokinetics: In vitro testing of small molecules, high throughput screening, MTT assay, survival curve, determination of IC50, synergy, cell proliferation, migration and death assays, cell cycle analysis using BrdU and 3HTdR incorporation, flow cytometry, immunohistochemistry, immunofluorescence.

Molecular Biology: Virus production and titer determination, transfection, transduction, cloning, DNA, RNA and protein isolation, histone extraction, PCR, gel electrophoresis, Southern blot, western blot, in situ hybridization.

Microtechnique: Perfusion of animals, fixation of tissue, cryostat and paraffin embedding, preparing sections using cryostat, microtome and vibratome.

Microscopy and Imaging: Light, DIC, confocal and fluorescence microscopy, time-lapse video-microscopy.

Mouse colony: Colony management, genotype analysis by PCR, retro-orbital bleed, tail-vein injection.

Computers: MS-Word, Excel, GraphPad Prism, Powerpoint, Adobe Photoshop, imaging softwares, statistical tools, use of internet.

Management: Managing projects to meet deadlines, independent judgement, coaching lab personnel, work equally well on self-managed projects or in a team environment, initiating and maintaining inter-lab relations.

EDUCATION

Doctor of Philosophy, Cell and Developmental Biology

“The Role of Presumptive Posterior Mesoderm in Morphogenesis and Cell Population Growth of Chick Embryo.”

University of Pune, India.

Master of Science, Zoology

University of Pune, India.

Bachelor of Science, Zoology

University of Pune, India.

PUBLICATIONS

1. Deshpande A.J., Chen, L., Bernt, K.M., Banka, D., Dias, S., Punt, N., and Armstrong, S.A.

Inhibition of MLL-AF9 mediated leukemogenesis by abrogating the DOT1L-AF10 interaction (manuscript in preparation).

2. Deshpande A.J., Fazio M., Chen, L., Bernt, K.M., Banka, D., Dias, S., Daigle, S., Richon V., Pollock, RM., Armstrong, S.A. Leukemic transformation by the MLL-AF6 Fusion Requires the H3K79 Methyltransferase Dot1l (manuscript in preparation).

3. Chen, Liying; Deshpande, Aniruddha,; Banka, Deepti ; Bernt, Kathrin M. ; Dias, Stuart; Buske, Christian; Daigle, Scott R; Richon, Victoria M; Pollock, Roy M; Armstrong, Scott A. Abrogation of MLL-AF10 and CALM-AF10 mediated transformation through genetic inactivation or pharmacological inhibition of the H3K79 methyltransferase Dot1l. (In revision).

4. Faust, P.L., Banka D., Siriratsivawong R, Ng V. G. and Wikander T. M. (2005). Peroxisome biogenesis disorders: The role of peroxisomes and metabolic dysfunction in the developing brain. J. Inherit. Metab. Dis. 28:369-383.

5. Sharmila Deshmukh-Barve, Deepti Joshi-Banka and Sohan P. Modak (2003). The proliferative state, graft site and contact-time of competent chick ectoblast determine the quantity of neural induction by Hensen’s node. Int. J Dev. Biol. 47(5): 355-361.

6. Deepti Joshi-Banka, Sharayu Paranjape, Surendra Ghaskadbi and Sohan P. Modak (2002). Post-nodal mesoblast caudalize host axis and inhibits cell population growth and induce new primitive streaks in chick embryo. Dev. Growth Differ. 44(2):169-180.

7. Deepti Joshi-Banka, Sharmila Deshmukh and Sohan P. Modak. (2001). The neural inductive signal is transferred to ectoblast in 1-2h but a continued contact with mesoblast for 2-3h is essential for neuralization in the chick area pellucida. Int. J. Dev. Biol. 45(5-6): 767-770.

ABSTRACTS AND PRESENTATIONS

• The comparison of various promoters for Wiskott Aldrich Syndrome gene therapy using insulated lentiviral vectors. American Society of Gene and Cell Therapy- 2010, Washington DC.

• Genetic influence on cerebellar growth and development: analysis of C57BL6/J and DBA/2J mice across embryonic and neonatal time points. Society for Neuroscience- 2006, Atlanta, GA.

• CNS intrinsic defects in developing peroxisome deficient cerebellar granule neurons. Society for Neuroscience- 2004, San Diego, CA.

• The inductive signal is transferred to ectoblast in 1-2 hours but a contact with mesoblast for additional 2-3 hours is essential for neural induction. NCBS Symposium on Cell and Developmental Biology- 2000, Bangalore, India.

• Postnodal fragment sustains the neural induction initiated by Hensen’s node in chick embryo. International Symposium on Development Growth and Differentiation- 1997, Mahabaleshwar, India.

• Postnodal mesoblast exerts negative cell cycle control and induces caudalization Indian Society of Developmental

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