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University of Pretoria

3 ARTICLES PUBLISHED IN JoVE

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 Creux^3,4, 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.

Genetics

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis

Andi M. Wilson^1,2, Brenda D. Wingfield^1,2

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.

Cancer Research

Flow Cytometric Analysis of Apoptotic Biomarkers in Actinomycin D-Treated SiHa Cervical Cancer Cells

Rivak Punchoo^1,2, 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.