Research Report 2003 - Max-Planck-Institut für molekulare Genetik

Department of Vertebrate Genomics
32
Oligofingerprinting / Cell Arrays Group
Scientists:
Dr. Anna Guerasimova
Dr. Dominique Vanhecke
Graduate students:
Andrea Fiebitz
Yuhui Hu
Scientific overview
The main research focus of the group comprises high-throughput functional genomics and
genome-wide expression analysis using transfected-cell array (TCA) and oligonucleotide fingerprinting
(ONF) technologies, respectively. In the field of functional genomics we optimised
and further developed a cell array platform based on the reverse transfection process (Figure 1).
Briefly, full-length open reading frames of genes inserted in expression vectors are printed at a
high density on a glass slide along with a lipid transfection reagent using a robotic arrayer.
Densities of up to 8000 spots per standard slide could be achieved. When the microarray of
DNA constructs are covered with a layer of adherent cells only the cells growing on top of the
DNA spots become transfected, resulting in the expression of specific proteins in spatially
distinctive groups of cells. The phenotypic effects
of this ‘reverse transfection’ of hundreds
or thousands of genes can be detected using
specific cell-based bioassays, e.g. immunofluorescence
(Figure 2) or induction of apoptosis.
At the moment we are focused on the development
of three applications using TCA:
Figure 1: Schematic presentation of the
principles of the transfected-cell array technique.
Head:
Dr. Michal Janitz
Fabeckstr. 60-62
14195 Berlin
Phone: +49 (0)30-8413 1486
Fax: +49 (0)30-8413 1462
Email: janitz@molgen.mpg.de
Technicians:
Nadine Scholz-Neumann
Irina Girnus
Sabine Thamm
Cellular localisation of the proteins
Taking advantage of transfection of hundreds
different cDNAs in parallel using the transfection
array we are able to determine
localisation of the expressed proteins in a highthroughput
manner using a microscope. By
evaluation of the transfected cells forming a
single spot we can determine whether the protein
expressed from the transfected cDNA
localises in the nucleus, cytoplasm or is transported
to the cellular membrane.