The team is studying the regulation of gene expression during the development of the immune system.
The main object of research is the NWC gene that we discovered, which is the third evolutionarily conserved gene in the RAG gene locus which are involved in the recombination of lymphocyte receptor genes. So far, we have learned about the mechanisms responsible for the mutual regulation of RAG and NWC gene expression during lymphocyte development, as well as the role that the NWC gene could play during the evolution of the RAG transposon in vertebrates. In addition, we showed that the NWC protein interacts with retrograde intraflagellar transport proteins and that its absence impairs sperm function in mice.
We are currently conducting research on the detailed definition of the role of NWC protein in intraflagellar transport and on the imaging of the impact of temporal silencing of NWC expression in Danio rerio on the ultrastructure of flagella and on the development of organs with flagellated cells. The second topic of research conducted by our team is the determination of the factors influencing the allele diversity of the sex determining gene (csd) in the honey bee population, the appropriate diversity of which is necessary for the viability of the population.
In the course of the conducted research, we found the presence of a much higher than previously expected number of csd alleles and their uneven distribution, indicating a very quick formation of new variants. Research is being conducted to determine the rate and mechanism of the formation of new alleles. Research on the genetic structure of the bee population is also aimed at determining its relationship with susceptibility to infectious diseases.
Laboratory Head
Head: Małgorzata Cebrat, PhD, associate professor (https://orcid.org/0000-0003-4544-3289)
MSc: biotechnology, Wroclaw University (1994-1999),
PhD: “Application of the cDNA-RDA technique in identifying differences in gene expression between normal and neoplastic cells of the small intestine of mice with the APC gene mutation”, Institute of Immunology and Experimental Therapy PAS (2002)
habilitation: „Function, regulation of expression and evolution of RAG/NWC locus genes”, Institute of Immunology and Experimental Therapy PAS (2010)
internships: Basel Institute for Immunology (2000-2001)
research interests: regulation of gene expression during the development of the immune system, evolutionary immunology, genetic structure of honey bee populations in the context of sex determination and resistance to infectious diseases
other: popularizing science (organizing and conducting molecular biology workshops for high school students), designing and making laboratory devices using 3D printing, beekeeping.
Team
Assistant Professors:
PhD Students:
- Robert Mroczek, MSc, PhD student
- Sylwia Janik, MSc, PhD student
Professors of the Institute:
- Paweł Kisielow, PhD, full professor
Most important scientific achievements
- discovery of the NWC gene, determination of factors involved in the control of NWC expression in the context of regulation of expression and evolution of the RAG gene locus playing a key role in lymphocyte development;
- demonstration of the role of the NWC protein in spermatogenesis;
- demonstrating the existence of a much larger than expected number of alleles of the gene responsible for sex determination in honey bees (csd) and their uneven distribution in the population.
Research methods
- analysis of gene expression and its regulation mechanisms (real-time PCR, analysis of promoter activity and epigenetic changes), sequencing;
- creation of genetic constructs using standard methods and homologous recombination, including large constructs (100-200 kb) and lentiviral vectors;
- confocal and fluorescence microscopy;
- analysis of protein expression and production of recombinant proteins in bacteria and mammalian cells;
- cell culture, transfections and transduction;
- study of interactions between proteins (fluorescence complementation, co-immunoprecipitation);
- transient silencing of gene expression in Danio rerio embryos and analysis of their phenotype;
- establishment of transgenic mice lines, Danio rerio breeding, bee breeding.
Key equipment
- ABI PRISM 310 Genetic Analyzer capillary sequencer
- molecular biology equipment
- cell biology equipment
- FDM and SLA/DLP 3D printers
Most important projects (during the last 10 years)
- „Factors influencing the diversity of csd alleles in honey bee (Apis mellifera) population”, National Science Centre, OPUS, 2019-2022, Małgorzata Cebrat
- „Role of NWC protein in the structure and functioning of cilia in Danio rerio”, National Science Centre, MINIATURA, 2020-2021, Joanna Niedbalska-Tarnowska
- „The role of bidirectional activity of the promoter of NWC gene in the evolution of RAG transposon”, National Science Centre. PRELUDIUM, Łukasz Śnieżewski
- „Mechanisms controlling DNA methylation in RAG/NWC locus”, Ministry of Science and Higher Education, 2010 – 2013, Małgorzata Cebrat
- „Production and characterization of genetically modified mice with a conditional knockout of NWC protein”, Ministry of Science and Higher Education, 2010 – 2013, Paweł Kisielow
Selected publications
- “Identification of a third evolutionarily conserved gene within the RAG locus and its RAG1-dependent and -independent regulation”; Cebrat M, Miazek A, Kisielow P. Eur J Immunol. 2005 Jul;35(7):2230-8. doi: 10.1002/eji.200526225. PMID: 15971274
- “Bidirectional activity of the NWC promoter is responsible for RAG-2 transcription in non-lymphoid cells”; Laszkiewicz A, Sniezewski L, Kasztura M, Bzdzion L, Cebrat M, Kisielow P. PLoS One. 2012;7(9):e44807. doi: 10.1371/journal.pone.0044807. Epub 2012 Sep 11. PMID: 22984564
- “Ikaros and RAG-2-mediated antisense transcription are responsible for lymphocyte-specific inactivation of NWC promoter”; Laszkiewicz A, Bzdzion Ł, Kasztura M, Snieżewski L, Janik S, Kisielow P, Cebrat M. PLoS One. 2014 Sep 8;9(9):e106927. doi: 10.1371/journal.pone.0106927. eCollection 2014. PMID: 25198102
- “Search for the Function of NWC, Third Gene Within RAG Locus: Generation and Characterization of NWC-Deficient Mice”; Kasztura M, Sniezewski L, Laszkiewicz A, Majkowski M, Kobak K, Peczek K, Janik S, Kapusniak V, Miazek A, Cebrat M, Kisielow P. Arch Immunol Ther Exp (Warsz). 2016 Aug;64(4):311-9. doi: 10.1007/s00005-015-0379-1. Epub 2015 Dec 24. PMID: 26703212
- “The evolutionary conservation of the bidirectional activity of the NWC gene promoter in jawed vertebrates and the domestication of the RAG transposon”; Sniezewski L, Janik S, Laszkiewicz A, Majkowski M, Kisielow P, Cebrat M. Dev Comp Immunol. 2018 Apr;81:105-115. doi: 10.1016/j.dci.2017.11.013. Epub 2017 Nov 22. PMID: 29175053
- „Lack of NWC protein (c11orf74 homolog) in murine spermatogenesis results in reduced sperm competitiveness and impaired ability to fertilize egg cells in vitro”; Majkowski M, Laszkiewicz A, Sniezewski L, Grzmil P, Pawlicka B, Tomczyk I, Michniewicz M, Kapusniak V, Janik S, Chodaczek G, Cebrat M. PLoS One. 2018 Dec 6;13(12):e0208649. doi: 10.1371/journal.pone.0208649. eCollection 2018. PMID: 30521625
- “Regulation of vitamin D receptor expression by retinoic acid receptor alpha in acute myeloid leukemia cells”; Marchwicka A, Cebrat M, Łaszkiewicz A, Śnieżewski Ł, Brown G, Marcinkowska E. J Steroid Biochem Mol Biol. 2016 May;159:121-30. doi: 10.1016/j.jsbmb.2016.03.013. Epub 2016 Mar 8. PMID: 26969398
- “Diverse Regulation of Vitamin D Receptor Gene Expression by 1,25-Dihydroxyvitamin D and ATRA in Murine and Human Blood Cells at Early Stages of Their Differentiation”; Janik S, Nowak U, Łaszkiewicz A, Satyr A, Majkowski M, Marchwicka A, Śnieżewski Ł, Berkowska K, Gabryś M, Cebrat M, Marcinkowska E. Int J Mol Sci. 2017 Jun 21;18(6):1323. doi: 10.3390/ijms18061323. PMID: 28635660
- “Microsporidia Nosema spp. – obligate bee parasites are transmitted by air”; Sulborska A, Horecka B, Cebrat M, Kowalczyk M, Skrzypek TH, Kazimierczak W, Trytek M, Borsuk G. Sci Rep. 2019 Oct 7;9(1):14376. doi: 10.1038/s41598-019-50974-8. PMID: 31591451
- “Uneven distribution of complementary sex determiner (csd) alleles in Apis mellifera population”; Zareba J, Blazej P, Laszkiewicz A, Sniezewski L, Majkowski M, Janik S, Cebrat M. Sci Rep. 2017 May 24;7(1):2317. doi: 10.1038/s41598-017-02629-9. PMID: 28539589
