Princeton University Office of Technology Licensing

Fourth Floor, New South Building

Post Office Box 36

Princeton, New Jersey 08544-0036

Phone: (609) 258-1570

FAX: (609) 258-1159

METHOD FOR ANALYSIS OF GENETIC INTERACTIONS

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