Prof. Maja Cemazar was a supervisor of PhD thesis of young researcher Tanja Dolinsek. In 2014, Tanja Dolinsek successfully defended her PhD thesis entiteld Antiangiogenic and antitumor effect of gene therapy with short noncoding RNA molecules against endoglin in murine tumor models.
D.09 Tutoring for postgraduate students
COBISS.SI-ID: 31328729In the invited lecture a method of non-invasive fluorescence imaging and its usability in gene therapy experiments was presented from the animal welfare point of view. In vivo imaging techniques represent exciting opportunity to conduct noninvasive and longitudinal log-term studies of dynamic biological processes. In gene therapy experiments non-invasive fluorescence imaging can be employed for following transgene expression in vivo and indirectly also the activity of promoters that drive their expression. In the lecture an example of CMV promoter activity study was used to show how the method can be used in gene therapy experiments. In the mentioned study we demonstrated that non-invasive fluorescence imaging is indeed an appropriate and convenient method to monitor the activity of the promoter in vivo which also promotes the principal ethical code of animal experiments, that is of 3Rs (replacement, reduction refinement).
B.04 Guest lecture
COBISS.SI-ID: 1811067In comparison to gene delivery with viral vectors »nonviral« gene delivery approaches offer several advantages, including safety, lower toxicity and easier preparation. Selective marker genes coding for antibiotic resistance have been an essential part of therapeutic plasmids, allowing selection of recombinant plasmids and retention in the bacterial host strain during large scale amplification. Due to the global spread of antibiotic resistant bacteria in the last two decades which makes it difficult to treat certain human and animal infections, health authorities recommend the ban of resistance genes for the construction of gene therapy vectors in order to prevent horizontal gene transfer of the resistance genes. The aim of the study was (1) to find out whether the ampicilin resistance gene from plasmid pORF-hIL12, which was used for electrogene therapy of mast cell tumors (MCT) in dogs, could be detected two and seven days after the treatment in the dog skin microbiota and (2) survey of dog skin microbiota before treatment in order to estimate the transformation efficiency of identified bacterial species with pORF-hIL12 in vitro. The study included 6 dogs with cutaneous or subcutaneous MCT, treated with intratumoral EGT. Swab samples taken from dog skin immediately before gene therapy and after 2 and 7 days were cultured on standard media. DNA was extracted from bacterial cells by the boiling metod or with DNA extraction kits. The ampiciline resistance gene and/or hIL12 gene were detected by PCR using different primers and protocols. Electroporation according to bacterial species protocols, where available, was used for in vitro transformation experiments. In none of the six treated patients, bacteria containing ampicilin or any other gene from plasmid pORF-hIL12 were detected after culturing. In vitro transformation of pORF-hIL12 into untreated and competent bacteria isolated from dog skin, was unsuccessful with the majority of the tested isolates. Only competent E.coli and Klebsiella sp. strains could be transformed with pORF-hIL12 at a high frequency in vitro. Our results demonstrate that horizontal transfer of the ampicilin resistance gene from pORF-hIL12 into bacteria present on dog skin is rather unlikely due to failure of successful transformation and/or replication of the plasmid in the recipient bacterial cells.
B.04 Guest lecture
COBISS.SI-ID: 3667066Prof. Maja Cemazar is editor of international journal Radiology and Oncology. Radiology and Oncology is a multidisciplinary journal promoting Slovenian and international research in the field of radiology and oncology internationally.
C.04 Editorial board of an international magazine