Sagar Mahale

I possess diverse expertise spanning Cell and Molecular biology, RNA biology, Pharmacy, and Pharmacology. My skills encompass various in-vitro cell biological assays development, including siRNA, LNA-ASO, and CRISPR-based tools, as well as advanced technologies like high-resolution, high content, multiplex microscopy. I have experience with a wide range of cell lines, including primary cells and cancer lines.

Ultimately, I aspire to leverage my pharmacological background to contribute to innovative drug discovery, translating molecular insights into tangible advancements in human health. My enduring goal is to participate in the development of novel, safe and cost-effective therapeutic strategies for addressing human ailments, all while fostering collaboration and individual growth within the research environment.

• Cell biology and Molecular biology
• Mammalian cell culture (Human, Mouse, Rat, Monkey and Chinese hamster cell lines)
• Primary cells, Stem cell culture and differentiation
• SiRNA, shRNA (Inducible), ASO/LNA mediated knockdown
• CRISPR-Cas mediated gene editing (activation, inhibition, RNA tagging for imaging)
• Cell assays using Flow cytometry, FACS, ELISA
• Generation of cell lines stably expressing transgenes
• Generation of clonal cell lines (Using CRISPR and FACS)
• Lentiviral transduction of mammalian cells
• Chemical and metabolic synchronization of cells in different stages of cell cycle
• RNA preparations, isolation, RNA and protein half-life, stability or degradation assays
• Biochemical Assays:
• DNA and protein electrophoresis
• Western blotting
• Immunoprecipitation
• Affinity purification
• RNA pulldowns (In-vitro transcription)
• Chromatin immunoprecipitation (ChIP)
• Chromatin oligo-affinity precipitation (ChOP)
• Immunofluorescence assay (IF)
• Immunohistochemistry (IHC)
• Fluorescence in- situ hybridization (FISH)
• RNA-Scope in-situ (RNA-DNA-Protein codetection)
• Duo-Link protein-protein interaction IF assays
• Molecular biology:
• SgRNA cloning into 3rd generation CRISPR-vectors
• RT-PCR
• Cloning of cDNA into plasmid vectors
• Site directed mutagenesis
• Transcriptomics:
• RNA sequencing (NGS)-Data analysis (usegalaxy.org)
• Differential expression gene analysis (DEG’s) using usegalaxy.org
• GO Enrichment and pathway analysis
• Neurobiology:
• Dissections of mouse brains
• Culturing primary neurons
• Brain tissue IHC/IP
• Pharmacology:
• Pharmacological screening of herbal and synthetic drugs Bioassay: Quantal Assays Graded Response Assays
• Pharmacokinetic Study (Bioanalytical methods) Area under the concentration-time profiles (AUC) measurement Peak Plasma concentration measurement (Cmax)
• Conducting toxicity studies on animals (rats) Acute toxicity study Chronic toxicity study
• Conducting behavioral studies on animals
• Optical imaging:
• Multicolor-immunofluorescence imaging using fluorescence, confocal and super-resolution microscopes
• Time-lapse imaging using fluorescence, confocal microscopes, high-content imaging (CD7-auto)
• Qualitative and quantitative image analysis using Imaris, Image J, NIS-Elements, ZEN 2, and LAS Leica suit software’s
• Miscellaneous:
• Well trained in handling animals, Xenograft studies in mice and collection of samples for in-vivo studies
• Handling experience of various instruments used in pharmacological screening
• Experience in establishing and managing a functional laboratory from ground up during PhD

• Sagar Mahale, Meenakshi Setia, Bharat Prajapati, Santhilal Subhash, Mukesh Pratap Yadav, Subazini Thankaswamy Kosalai, Ananya Deshpande, Jagannath Kuchlyan, Mirco Di Marco, Fredrik Westerlund, L. Marcus Wilhelmsson, Meena Kanduri* and Chandrasekhar Kanduri*. HnRNPK maintains single strand RNA through controlling double-strand RNA in mammalian cells. Nature Communications, (2022), https://doi.org/10.1038/s41467-022-32537-0.
• Sagar Mahale, Meenakshi Setia, Silke Reischl, Timo Reitinger and Chandrasekhar Kanduri. Long noncoding RNA controlled monoubiquitination determines the functional switch of RSP3 between mitosis and interphase. Manuscript in preparation (2023).
• Samarjit Jana, Mainak Mondal, Sagar Mahale, Bhabna Gupta, Kaval Reddy Prasasvi, Lekha Kandasami, Neha Jha, Abhishek Chowdhury, Vani Santosh, Chandrasekhar Kanduri, Kumaravel Somasundaram. PITAR, a DNA damage-inducible Cancer/Testis long noncoding RNA, inactivates p53 by binding and stabilizing TRIM28 mRNA. Manuscript Under revision at eLife (2023).Amrita Kumari, Chandan Kumar, Rajaiah Pergu, Megha Kumar, Sagar Mahale, Neeraj Wasnik and Sivaram V S Mylavarapu. Phosphorylation and Pin1 binding to the LIC1 subunit selectively regulate mitotic dynein functions. Journal of cell biology, (2021) 6; 220(12): e202005184.
• Luisa Statello, Mohamad Moustafa Ali, Silke Reischl, Sagar Mahale, Subazini Thankaswamy Kosalai, Vijay Suresh Akhade, Maite Huarte and Chandrasekhar Kanduri. The DNA damage inducible lncRNA SCAT7 regulates genomic integrity and TOP1 turnover in lung adenocarcinoma. Nucleic Acids Research Cancer, (2021) Volume 3, Issue 1, https://doi.org/10.1093/narcan/zcab002
• Mohamad M. Ali, Mirco Di Marco, Sagar Mahale, Daniel Jachimowicz, Subazini Thankaswamy Kosalai, Silke Reischl, Luisa Statello, Kankadeb Mishra, Catarina Darfors, Meena Kanduri, Chandrasekhar Kanduri. LY6K-AS lncRNA is a lung adenocarcinoma prognostic biomarker and regulator of mitotic progression. Oncogene (2021). https://doi.org/10.1038/s41388-021-01696-7.
• Sagar P Mahale, Megha Kumar*, Amit Sharma *, Aswini Babu, Shashi Ranjan, Chetana Sachidanandan and Sivaram V S Mylavarapu. Dynein Light Intermediate Chain 2 Regulates Mitotic Spindle Orientation. Scientific Reports (2016), Article number: 22, doi:10.1038/s41598- 016-0030-3.
• Sagar P. Mahale, Amit Sharma and Sivaram V. S. Mylavarapu. Dynein Light Intermediate Chain 2 is required for inactivating the spindle assembly checkpoint at the metaphase to anaphase transition. PLoS One 11, (2016) e0159646, doi:10.1371/journal.pone.0159646.
• L.A. Pradeep Rajkumar, S. Vimal kumar, G.Nalini, T.Gayathri, Sagar Mahale, B.Lakshminarayanan. Evaluation of chronopharmacology and chronosensitivity of centrally acting potential CNS drugs in albino wistar rats. Annals of Biological Research, (2010), 1 (4): 52-56.
• M. K. Mishra, R. V Sahu, S. P. Mahale, M. P. Goojar, D.K. Dash, A. Rout, B. B. Barik. Antifungal potential of Phyllanthus niruri linn aerial parts. International Journal of Pharmacology and Biological Sciences (2010), Vol. 4 (2) p- 51-56.

• Gold Medal for the best scientific oral presentation at the AIST International Imaging Workshop at AIST, Tsukuba, Japan, Jan. 2014
• Best Scientific Poster award at RCB Colloquia, Gurgaon, India, Sept. 2014
• Summer Research Fellowship by Indian Academy Of Sciences (INSA), Bangalore, 2010

1. Name:       Sivaram Mylavarapu, PhD
   Position:   Professor at UNESCO-Regional Centre for Biotechnology (RCB)
   Email ID:   sivaram@rcb.res.in
   Phone:       +919650248636

2. Name:       Silke Reischl, PhD
   Position:    Senior scientist Bioscience at Astrazeneca, Göteborg, Sweden
   Email ID:   silke.reischl@web.de
   Phone:       +46734260166