University of Pretoria 3 articles published in JoVE Cancer Research Flow Cytometric Analysis of Apoptotic Biomarkers in Actinomycin D-Treated SiHa Cervical Cancer Cells Rivak Punchoo1,2, Esther Zhou1, Sachin Bhoora1 1Department of Chemical Pathology, Faculty of Health Sciences, University of Pretoria, 2Tshwane Academic Division, National Health Laboratory Service Apoptosis can be characterized by flow cytometric analysis of early and late apoptotic biomarkers. The cervical cancer cell line, SiHa, was analyzed for apoptosis biomarkers after treatment with Actinomycin D using a benchtop flow cytometer. Genetics CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis Andi M. Wilson1,2, Brenda D. Wingfield1,2 1Department of Biochemistry, Genetics & Microbiology, University of Pretoria, 2Forestry & Agricultural Biotechnology Institute (FABI), University of Pretoria The CRISPR-Cas9 genome editing system is an easy-to-use genome editor that has been used in model and non-model species. Here we present a protein-based version of this system that was used to introduce a premature stop codon into a mating gene of a non-model filamentous ascomycete fungus. Genetics The Use of Induced Somatic Sector Analysis (ISSA) for Studying Genes and Promoters Involved in Wood Formation and Secondary Stem Development Antanas Spokevicius1, Lynette Taylor1, Emma Melder1, Kim Van Beveren1, Josquin Tibbits2, Nicky Creux3,4, Gerd Bossinger1 1School of Ecosystem and Forest Sciences, Faculty of Science, The University of Melbourne, 2Victorian AgriBiosciences Centre, La Trobe University R&D Park, 3College of Biological Sciences, Department of Plant Biology, University of California, Davis, 4Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria Here we present a protocol that facilitates the medium to high throughput functional characterization of gene and promoter constructs in tree secondary stem tissue within comparatively short time frames. It is efficient, easy to use and widely applicable to a range of tree species.