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 GENETIC ANALYSIS OF ALTERNATIVE SPLICING

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

Publications continued:

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