Uma Muthukrishnan Email & Phone Number

Bengaluru, Ka, In

Bengaluru, Karnataka, India

Sweden

During my studies, I had gained in-depth knowledge and understanding of the extranuclear functions of histone proteins and exosome biogenesis. I have further acquired theoretical and practical skills in a range of techniques involving cell culture (primary and cell lines), cell-based assays, exosome isolation and purification using differential gradient methods, exosome uptake study, transfection, stable-cell line generation, CellineAD bioreactor maintenance, NTA (Nanoparticle Tracking Analysis) -based exosome characterization, immunocytochemistry, western blotting, etc.Developed skills and abilities for independent thought through planning, execution, and interpretation of the project work. I have learned to apply my own judgement to the design and direction of my work and also able to learn to prioritize the experiments/projects. I have also developed my supervisory and mentoring skills by supervising students working with me on the project.

Sweden

Setting up a platform for protein/exosome studies. Involved in the lab organisation.Isolation and maintenance of primary cell culture as well as rodent, murine, and human cell lines. Isolation of exosomes from mammalian cells (primary murine glial cells, rodent cell line) and examine proteins in SDS-PAGE and western blot. Proliferation assay on murine/rodent glial cells.

Bangalore

Project title: Identification of molecular targets of the transcription regulator ASCL1, which is differentially expressed in primary and secondary glioblastoma (GBM).Glioblastomas are the most common primary tumors of the brain. The tumors arising from the astrocytes (astrocytic tumors or astrocytoma) are histopathologically graded into four types according to their increasing malignancy. In order to find new diagnostic and therapeutic strategies, a better understanding of the biological pathway (s) leading to glial tumorigenesis is warranted. In one of the recent studies from the lab, ASCL1 (Achaete-scute complex homolog 1) has shown to be overexpressed in secondary GBM when compared with primary GBM. In addition to this, a significant reduction in ASCL1 protein via the Notch pathway leads to the suppression of tumor growth has been found in several cancers. My Goals in this project was Selection of suitable glial tumor cell line, which has a higher level of ASCL1 gene expression using quantitative real-time quantitative PCR method (qRT-PCR).Silencing of ASCL1 in the chosen glioma U87MG cell line using the siRNA approach.ASCL1 downregulation was quantified using qRT-PCR ( Eva Green (Biorad) on a Biorad iQ5 cycler ). Validated ASCL1 downregulation using semi-quantitative PCR and western blot. Total RNA samples from scrambled siRNA and siRNA-ASCL1 were hybridized onto Human Genome U133 Plus 2.0 GeneChip (Affymetrix) to perform microarray analysis. Designed primers for selected differentially expressed targets to validate them using ChIP-on-chip.Cloning of human ASCL1 cDNA in a mammalian expression vector to perform studies like immunofluoresence.

Bangalore

Supervisor: Prof. P. N. Rangarajan, Department of Biochemistry.Project 1:Analysis of one of the downregulated genes during neurotropic viral infection, Calcium Calmodulin Kinase II inhibitor alpha in different mammalian species. Techniques: SDS-PAGE, Western blot, Lysate preparationProject 2:Cloning and expression of Mxr1p, a transcription factor involved in Alcohol oxidase gene regulation in Pichia pastoris.Cloning, SDS-PAGE, 2D-gel electrophoresis, FPLC (using AKTA explorer), western blot

Licentiate Degree, Molecular Medicine

Master'S Degree, Genomics

Bachelor'S Degree, Biotechnology