Molecular Biology Laboratory

Department of Biology and Pharmaceutical Botany

Faculty of Pharmacy

OUTLINE OF THE LABORATORY

The unit’s research activities focus on using molecular biology techniques with particular emphasis on transcriptomic studies. We are conducting expression analyses of genes involved in the unfolded protein response (UPR) pathway. In addition, using the foregoing method, we also determine changes in the miRNA levels with documented regulatory roles in the course of the UPR. Basic research, which is the goal of our scientific activity, allows us to expand knowledge regarding the mechanism of the UPR, whose disorders contribute to the development of numerous diseases, i.e. diabetes, neurodegenerative disorders, or cancer. A deeper understanding of the stress response enables further exploration of the application aspects, which may result in the development of new therapeutic strategies in the future.

We conduct cell cultures using both primary and tumor lines for experiments. Our cultures are regularly monitored for Mycoplasma sp. contamination. We isolate RNA and a fraction of miRNA from the specimens collected to ensure further analysis. Our tests are performed at both the RNA and protein levels (using Western blot). Nuclear and cytosolic fractions are isolated with a high degree of purity using the Paris Kit.

We have extensive experience in cell transfection using lipofectamine. Our activities involve using siRNAs, miRNA analogs, and their inhibitors, target protector molecules. Satisfactory silencing and overexpression are achieved with little cytotoxic effect.

AVAILABLE EQUIPMENT AND APPARATUS

We have the equipment necessary for Real-Time Cell Analysis. Results are validated by examining the activity of major executive caspases (Caspase Glo 3/7 Assay). With the ability to use luminometric methods, we also conduct research using reporter genes (Dual-Luciferase Reporter Assay).

PUBLICATIONS

Publications that have been developed by using the research methods described:

1. Gebert M, Bartoszewska S, Janaszak-Jasiecka A, Moszyńska A, Cabaj A, Króliczewski J, Madanecki P, Ochocka RJ, Crossman DK, Collawn JF, Bartoszewski R. PIWI proteins contribute to apoptosis during the UPR in human airway epithelial cells. Sci Rep. 2018 Nov 6;8(1):16431. doi: 10.1038/s41598-018-34861-2. PMID: 30401887; PMCID: PMC6219583

2. Bartoszewski R, Gebert M, Janaszak-Jasiecka A, Cabaj A, Króliczewski J, Bartoszewska S, Sobolewska A, Crossman DK, Ochocka R, Kamysz W, Kalinowski L, Dąbrowski M, Collawn JF. Genome-wide mRNA profiling identifies RCAN1 and GADD45A as regulators of the transitional switch from survival to apoptosis during ER stress. FEBS J. 2020 Jul;287(14):2923-2947. doi: 10.1111/febs.15195. Epub 2020 Jan 10. PMID: 31880863

3. Gebert M, Sobolewska A, Bartoszewska S, Cabaj A, Crossman DK, Króliczewski J, Madanecki P, Dąbrowski M, Collawn JF, Bartoszewski R. Genome-wide mRNA profiling identifies X-box-binding protein 1 (XBP1) as an IRE1 and PUMA repressor. Cell Mol Life Sci. 2021 Nov;78(21-22):7061-7080. doi: 10.1007/s00018-021-03952-1. Epub 2021 Oct 12. PMID: 34636989; PMCID: PMC8558229

4. Bartoszewska S, Cabaj A, Dąbrowski M, Collawn JF, Bartoszewski R. miR-34c-5p modulates X-box-binding protein 1 (XBP1) expression during the adaptive phase of the unfolded protein response. FASEB J. 2019 Oct;33(10):11541-11554. doi: 10.1096/fj.201900600RR. Epub 2019 Jul 17. PMID: 31314593; PMCID: PMC6766644

Price list:

The cost of services is subject to individual pricing.

CONTACT

Magdalena Gebert, MPharm.
magdalena.gebert@gumed.edu.pl

Molecular Biology Laboratory
Department of Biology and Pharmaceutical Botany
Faculty of Pharmacy
Medical University of Gdańsk
Al. Gen. J. Hallera 107
80-416 Gdańsk

phone 58 349 13 99