Our laboratory studies
how naturally occurring transcription factors mediate
specific interactions with DNA and other proteins
(molecular recognition) and uses the general principles
derived from this work to construct synthetic “designer”
transcription factors (protein engineering).
Our efforts focus primarily on Cys2His2 zinc fingers,
the most common structural motif found in eukaryotic
transcription factors. Cys2His2 zinc finger proteins
mediate specific protein-DNA and protein-protein interactions,
many of which are central to normal and disease-related
cell processes. In addition, selection and design
methods have been used to engineer synthetic Cys2His2
zinc fingers capable of recognizing novel DNA sequences
of interest.
Structure-function studies of biologically important
zinc finger proteins
We use a combination of genetic, biochemical, structural,
and mammalian cell-based approaches to gain a physical-chemical
understanding of DNA and protein recognition mediated
by biologically important zinc finger proteins. Two
major projects in the lab focus on studying the recognition
of extended DNA sequences by the NRSF/ REST zinc finger
repressor and on understanding how zinc fingers in
the Ikaros family of transcription factors mediate
specific homo- and hetero-typic interactions.
Engineering “designer” zinc fingers
with novel DNA-binding specificities
Using high-throughput selection and design methods
developed in our lab, we are compiling “libraries”
of synthetic zinc finger domains with novel, defined
DNA binding specificities. These “designer”
zinc fingers provide powerful tools for biological
research and potentially for gene therapy as they
can be used to target functional domains to any desired
gene of interest. For example, designer zinc fingers
fused to a non-specific DNA cleavage domain can be
used to introduce double-stranded DNA breaks into
specific genomic loci in a mammalian cell. The site-specific
breaks introduced by such “zinc finger nucleases”
can be used to promote homologous recombination and
gene targeting at specific genomic loci.
|
