Sahithi Seedarala

• Stable cell line generation - retrovirus and lentivirus systems • Small and large-scale DNA and RNA isolation • Western blot, immunofluorescence, co-IP, viability assay, migration assay • Primer designing, PCR-based cloning, Gibson assembly • Primary melanocytes isolation from mice • DNA amplicon libraries generation for NGS sequencing

• Small and large-scale bacterial (E. coli), mammalian (HEK-293, Expi-HEK, CHO, Expi-CHO), and baculovirus (Tni and Sf9) protein expression up to 10L • Small-scale Pro-A and His-tag-based protein purification • Protein quality control- SDS-PAGE, Western Blot • Co-ordination with project managers and protein purification team

• Patient biopsy-derived cell line maintenance -2D and 3D cell culture • HTS immunofluorescence assay and cell viability assay development to understand drug resistance in cancer patients

• Develop 2D SDS-PAGE, Western blot to analyze Host Cell Proteins (HCP) in Biosimilar • Small scale Protein-A based affinity purification, CHO-HCP ELISA • GMP-compliant laboratory

• Evaluation of inhibitory action of novel MAPK inhibitors and HSP90 inhibitors in various cell lines by MTS Anti-proliferation assays and Western Blot

• Expressed and purified S.aureus and M. smegmatis DNA Gyrase A and B proteins in E.coli • Performed DNA supercoiling assays to evaluate the effectiveness of novel DNA Gyrase B inhibitors • Performed high-throughput screening of Chorismate mutase inhibitors by cell-based Microplate Alamar Blue assays (MABA) and Chorismate Mutase enzyme assays

Gq/11-mediated GPCR signaling in melanoma pathogenesis and therapy. Understanding the transactivation mechanism between GPCR and Hgf/c-MEt signaling in melanoma