Laboratory of Cell Engineering
Head: Agata KLEJMAN
Staff: Joanna CHILCZUK, Paulina KOZA
The Laboratory of Cell Engineering was established at the Nencki Institute of Experimental Biology in March 2011. The facility is a fully equipped professional laboratory specialized in the production of genetically modified animals as well as viral vectors. The Laboratory provides the core facility service for investigators from the Nencki Institute and other scientific and R&D institutions.
Genetically modified animals (mice and rats) are produced by use of classical and lentiviral transgenesis methods. We possess high expertise in producing transgenic rats, which compared to mice, represent far more useful in vivo models to study mechanisms of human diseases. We are also introducing the “knock-out” technology into our Laboratory.
We have substantial experience in obtaining lentiviral vectors with tissue-specific (mainly neurospecific) expression pattern as well as the shRNA expressing vectors. Lentiviruses are currently extensively used in scientific and medical approach, as they constitute the most efficient tool able to incorporate transgene of interest into genome of non-dividing as well as dividing cells
More information about laboratory on the web page: http://celleng.nencki.gov.pl/ and lce@nencki.gov.pl
Genetically Modified Animal Models:
- Preparing of constructs (choosing the promoter, cloning, confirming the expression by in vitro assays)
- Microinjection of zygotes
- Traditional method (injecting of DNA solution into one of the pronuclei of fertilized embryo)
- Using the lentiviral vectors (incorporating of viruses under the zona pellucida) - Oviduct transfer of the injected zygotes into pregnant/pseudopregnant females
- Vasectomy in males
- Tail or ear tissue biopsy, DNA isolation
- Genotyping
Viral vectors technology:
- Expertise and technical assistance regarding choosing of the tissue specific promoters, projecting and producing vectors carrying silencing sequences
- Cloning, preparing the plasmids for production of viruses
- Production of the viral vectors (lenti, adenoviral)
- Titer analysis (Real-Time PCR)
- Further experiments with created viral vectors (e.g. developing of stable cell lines by use of lentiviral vectors)
Selected publications:
Konopka W., Duniec K., Klejman A., Wawrzyniak W., Owczarek D., Gawryś L., Maleszewski M., Mallet J., Kaczmarek L. (2009) Tet system in the brain: transgenic rats and lentiviral vectors approach. Genesis, 47: 274-80.
Uckermann O., Luksch H., Stefovska V., Hoehna Y., Marzahn J., Theil M., Pesic M., Górkiewicz T., Gawlak M., Wilczyński G.M., Kaczmarek L., Ikonomidou C. (2011) Matrix metalloprotein- ases 2 and 9 fail to influence drug-induced neuroapoptosis in developing rat brain. Neurotoxicity Research, 19: 638-48.
Wilczyński G.M., Konopacki F.A., Wilczek E., Lasiecka Z., Gorlewicz A., Michaluk P., Wawrzyniak M., Malinowska M., Okulski P., Kołodziej L.R., Konopka W., Duniec K., Miodu- szewska B., Nikolaev E., Walczak A., Owczarek D., Górecki D.C., Zuschratter W., Ottersen O.P., Kaczmarek L. (2008) Important role of matrix metalloproteinase 9 in epileptogenesis. Journal of Cell Biology, 180: 1021-35.
