Irell & Manella Graduate School of Biological Sciences

3 ARTICLES PUBLISHED IN JoVE

Biology

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Lauren Liddell^1,2, Glenn Manthey², Nicholas Pannunzio³, Adam Bailis²

¹Irell & Manella Graduate School of Biological Sciences, ²Department of Molecular and Cellular Biology, City of Hope Comprehensive Cancer Center and Beckman Research Institute, ³Department of Biochemistry and Molecular Biology, University of Southern California, Norris Comprehensive Cancer Center

The HO-stimulated translocation assay monitors single-strand annealing following the creation of DNA double-strand breaks at multiple loci in diploid Saccharomyces cerevisiae. This mechanism may model genome rearrangements in somatic cells of higher eukaryotes following exposure to high doses of ionizing radiation.

Immunology and Infection

Humanized NOD/SCID/IL2rγ^null (hu-NSG) Mouse Model for HIV Replication and Latency Studies

Xin Xia¹, Haitang Li¹, Sangeetha Satheesan^1,2, Jiehua Zhou¹, John J. Rossi¹

¹Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, ²Irell and Manela Graduate School of Biological Sciences, Beckman Research Institute of City of Hope

This protocol provides a method to establish humanized mice (hu-NSG) via intrahepatic injection of human hematopoietic stem cells into radiation-conditioned neonatal NSG mice. The hu-NSG mouse is susceptible to HIV infection and combinatorial antiretroviral therapy (cART) and serves as a suitable pathophysiological model for HIV replication and latency investigations.

Immunology and Infection

Identification of Nucleolar Factors During HIV-1 Replication Through Rev Immunoprecipitation and Mass Spectrometry

Jerlisa Ann C. Arizala^1,2, Pritsana Chomchan¹, Haitang Li¹, Roger Moore³, Helen Ge³, Dominique L. Ouellet¹, John J. Rossi^1,2

¹Molecular and Cellular Biology Department, Beckman Research Institute at the City of Hope, ²Irell & Manella Graduate School of Biological Sciences, ³Department of Molecular Immunology, Beckman Research Institute and the City of Hope

Here we describe Rev immunoprecipitation in the presence of HIV-1 replication for mass spectrometry. The methods described can be used for the identification of nucleolar factors involved in the HIV-1 infectious cycle and are applicable to other disease models for the characterization of understudied pathways.