3 articles published in JoVE
Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs Sabyasachi Dash1,2,3, Muthukumar Balasubramaniam2, Chandravanu Dash2, Jui Pandhare2 1School of Biotechnology, Kalinga Institute of Industrial Technology University, 21. Center for AIDS Health Disparities Research Department of Biochemistry and Cancer Biology, Meharry Medical College, 3Kalinga Institute of Industrial Technology University This report describes a fast and reliable method for validating mRNA targets of cellular miRNAs. The method uses synthetic biotinylated Locked Nucleic Acid (LNA)-based miRNA mimics to capture target mRNA. Subsequently, streptavidin-coated magnetic beads are employed to pulldown the target mRNA for quantification by qPCR polymerase chain reaction.
Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes Muthukumar Balasubramaniam1,2, Benem Davids1,2, Amma B. Addai1,2, Jui Pandhare1,3,4, Chandravanu Dash1,2,5 1Center for AIDS Health Disparities Research, Meharry Medical College, 2Department of Biochemistry and Cancer Biology, Meharry Medical College, 3School of Graduate Studies and Research, Meharry Medical College, 4Department of Microbiology and Immunology, Meharry Medical College, 5Tennessee Center for AIDS Research (TN-CFAR), Meharry Medical College This report describes an in vitro assay for measuring the human immunodeficiency virus type 1 (HIV-1) preintegration complex (PIC)-associated integration activity in the cytoplasmic extracts of acutely infected cells. The integration of PIC-associated HIV-1 DNA into heterologous target DNA is quantified by using a nested real-time polymerase chain reaction strategy.
Identifying Dysregulated Genes Induced by Kaposi's Sarcoma-associated Herpesvirus (KSHV) Donald Alcendor1, Susan Knobel1 1Department of Microbiology & Immunology and the Center for AIDS Health Disparities Research, Meharry Medical College Host cell factors play a critical role in the establishment and maintenance of Kaposi's sarcoma (KS). We outline methods to identify host cell factors altered in KSHV-infected DMVEC cells, and in KS tumor tissue. Cellular genes altered by virus will serve as potential target(s) for novel therapeutics.