Description:
Researchers at Princeton University have developed methods useful in
large-scale probing of interactions between genes and their products. More
specifically, the new technology is useful for a systematic parallel analysis of
genetic and protein-protein interactions on a genome-wide scale. For example,
the developed technology can be applied for massive detection of gene
combinations that promote generation of cancer or induce stem cell
differentiation toward specific lineages. Princeton is currently seeking
industrial collaborators to commercialize this technology.
Identification of physical and functional interactions between gene
products is highly important for analysis of multifaceted biological disorders
such as cancer. For example, to undergo cancer transformation, a biological cell
often needs to accumulate a series of genetic changes that promote simultaneous
misexpression in more than one gene. To analyze functional consequences of
simultaneous gene misexpression, the pairs of DNAs encoding different genes
(cDNAs) are introduced simultaneously into cells of interest. If a pair of genes
promotes uncontrolled cell division, as compared to each of these genes tested
separately, it can be classified as synergetic genetic interaction. Given that
number of genes in human is more than 20,000, the estimated number of gene pairs
to test is more than 100,000,000. If one would spend only 1 hour for analysis of
each gene pair, it would take more than 10,000 years to identify all the
possible gene-gene interactions in a single cell type!
To facilitate large-scale identification of genetic interaction, the
Princeton researchers suggest a microarray-based method of parallel analysis. In
their approach, pairs of cDNAs (or RNAi) from different genes are placed on a
same vector, virus or plasmid-based. Each vector molecule has a unique
identifier, a ¿bar-code¿ sequence, which is also represented on microarrays. A
library of the ¿bar-coded¿ cDNA pairs is introduced into cells of interest that
are further propagated at selective conditions. A cell will survive if it
includes a functional combination of cDNAs. In this way, combinations of genetic
elements with synergetic action undergo genetic selection for their ability to
maintain a specific cellular phenotype. At the end of the screen, the vector DNA
is extracted from cells and changes in representation of each ¿bar-code¿ are
measured using microarrays. This approach allows detecting functionally
synergetic pairs of genes.
The Princeton approach provides a highly cost-effective method for
analysis of the genetic interactions. For example, this technology can be used
to identify specific combinations of cDNAs or RNAis that promote uncontrolled
cell growth in tumors or induce spontaneous cell death. It can be easily 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 collaborators to commercialize the is 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, combination, interaction, bar-code.
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