Chemically synthesized zinc finger molecules as nano-addressable probes for double-stranded DNAs.
Our experiments describe an alternative method of dsDNA recognition using zinc finger (ZF) molecules which bind DNA specifically and with high affinity. Our aim was to develop zinc finger probes which are able to bind to dsDNA molecules at predetermined sites. In our basic approach we used pairs of complementary oligonucleotides to form dsDNAs, containing one of the three SP1-transcription factor motifs as a zinc finger recognition site. Two zinc finger probes of the SP1 motif were chemically synthesized and modified with a Dy-633 fluorophore. The SP1 peptides were folded into functional zinc fingers using zinc chloride. The addressable dsDNAs were immobilized on optical fibres, and the kinetics and binding rates of the artificial zinc finger probes were measured by a fluorescence detecting device (photomultiplying tube). The two zinc fingers and their corresponding DNA recognition sites served as specific probes and controls for the matching site and vice versa. Our experiments showed that a variety of dsDNA-binding probes may be created by modification of the amino acid sequence of natural zinc finger proteins. Our findings offer an alternative approach of addressing dsDNA molecules, for example for use in a nanoarray device.
von Nickisch-Rosenegk
Markus
M
Dpt. Molecular Bioanalytics and Bioelectronics, Fraunhofer Institute of Biomedical Engineering (IBMT), Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany. markus.nickisch@ibmt.fhg.de
Ehrentreich-Forster
Eva
E
Strehlow
Rothin
R
Christmann
Alexander
A
Bier
Frank F
FF