Description:
Researchers at
Princeton University have developed methods useful in genetic high-throughput
screens (HTS) of alternative splice variants. More specifically, the developed
technology can be applied for massive functional identification of splice
variants whose misexpression can promote cancerous cell transformation or induce
stem cell differentiation toward specific lineages. Princeton is currently
seeking industrial collaborators to commercialize this
technology.
More than 70% of
human genes undergo alternative splicing, a process that assemble different
combinations of exons to produce mRNA isoforms with distinct protein coding
potentials. However, the biological functions for the vast majority of the
splice variants remain unknown. To address this problem, Princeton researchers
have developed a method that utilizes special exon arrays for a massive parallel
functional analysis of splice variants in the following fashion. A complex cDNA
library carried on a plasmid or virus vector is introduced into cells of
interest. The cells are propagated at selective (harsh) conditions. In such
conditions, a cell will survive if it includes a functional cDNA representing a
particular splice variant. After the selection, the vector DNA is extracted from
the cells, labeled and hybridized to exon microarrays. The subsequent analysis
of the microarray data allows identifying splice variants who rescue cell growth
at the selective conditions.
The described
approach provides a highly cost-effective method for large-scale identification
of splice variants that function to produce a specific cell phenotype. For
example, this technology can be used to identify splice variants that promote
uncontrolled cell growth in tumors. It can be adapted to various research models
in vitro and in vivo and provide a unique platform
for drug discovery and development of novel therapies.
Princeton is currently seeking
industrial collaboration to commercialize this technology. Patent protection is
pending.
Publications:
Diversification of Stem Cell Molecular
Repertoire by Alternative Splicing, Pritsker et al. 2005 PNAS, Vol 102,
No.40
Genomewide gain-of-function genetic
screen identifies functionally active genes in mouse embryonic stem cells.
Pritsker et al. 2006 PNAS, Vol. 103,
No.18
Alternative Splicing Increases
Complexity of Stem Cell Transcriptome, Cell Cycle 5:4, 347-351, 16, February
2006
Keywords:
HTS, microarray, genetic screen,
library, phenotype, function, genome, cDNA, RNAi, overexpression, tumor, cancer,
stem cells, alternative splicing, exon.
For more information please contact:
Laurie Tzodikov
Office of Technology Licensing
Princeton University
4 New South Building
Princeton, NJ 08544-0036
(609) 258-7256
(609) 258-1159 fax
tzodikov@princeton.edu
