Meharry Medical College

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Identifying Dysregulated Genes Induced by Kaposi's Sarcoma-associated Herpesvirus (KSHV)

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.

Immunology and Infection

Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes

Muthukumar Balasubramaniam^1,2, Benem Davids^1,2, Amma B. Addai^1,2, Jui Pandhare^1,3,4, Chandravanu Dash^1,2,5

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.

Genetics

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs

Sabyasachi Dash^1,2,3, 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.