Princeton Docket # 11-2672

Researchers at Princeton University have developed a novel system for the rapid and specific induction of individual genes, and upon modification, this system also allows for the rapid and specific degradation of target proteins.  Substantial induction/degradation of target genes/proteins can be achieved within minutes following the addition of an inducing molecule, b-estradiol.  This inducer was shown to be truly gratuitous in that it interacts with no other regulatory system and has minimal effects on the physiology of the cells except for target-specific degradation and downstream effects.  Although demonstrated in yeast, this system is potentially applicable to other eukaryotes.  This novel system has potential applications in synthetic biology, pharmaceutical and agricultural industries, and in basic biomedical research.

Current methods for controlling protein abundance in yeast include utilization of tetracycline-inducible tetR-based systems, the GAL4-UAS system, and the chimeric GEV system. Each of these systems,  in their current form,  has significant shortcomings. Tetracycline-based systems can turn a target gene on or off at the transcriptional level, but cannot exact control at the protein level.  As many proteins have long half-lives, the tetracycline-based systems are not efficient in depleting target proteins.  Moreover, the GAL4-UAS system results in deleterious physiological changes because it requires that cells be grown in relatively poor, non-glucose carbon sources prior to an inducing galactose pulse.  Lastly, previous iterations of the GEV system have either relied on plasmids or have been only partially characterized. The GEV system has only been devised to induce gene expression, but not protein depletion.

Applications            

·         Target identification

·         Bioengineering for biologic/compound synthesis

·         High throughput screening of compound libraries

·         Research tool to study gene function

Advantages  

·         Induction/degradation achieved within minutes

·         Highly specific control over target proteins

·         Unchanged cellular physiology

·         Gratuitous inducer

 Publications

McIsaac RS, Silverman SJ, McClean MN, Gibney PA, Macinskas J, Hickman MJ, Petti AA, Botstein D.  Fast-acting and nearly gratuitous induction of gene expression and protein depletion in Saccharomyces cerevisiae. Mol Biol Cell. 2011 Sep 30.

Hickman MJ, Petti AA, Ho-Shing O, Silverman SJ, McIsaac RS, Lee TA, Botstein D. Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast. Mol Biol Cell. 2011 Sep 7.

Inventors

David Botstein is Anthony B. Evnin Professor of Genomics and Director of the Lewis-Sigler Institute for Integrative Genomics at Princeton University.  His research areas includes: (1) genome-wide studies of gene expression through the life cycle and experimental evolution of budding yeast (Saccharomyces cerevisiae), (2) mechanisms by which yeast maintain metabolic homeostasis in the face of environmental and genetic perturbations, and (3) quantitative analysis and intuitive display of genome-scale biological information in the context of genomic databases.

Professor Botstein graduated from Harvard in 1963 and received a Ph.D. from the University of Michigan in 1967.  He then taught at the Massachusetts Institute of Technology, where he became a Professor of Genetics.  Dr. Botstein joined Genentech, Inc. in 1987 as Vice President-Science.  In 1990, he became Chairman of the Department of Genetics at Stanford University.  Dr. Botstein was elected to the U.S. National Academy of Sciences in 1981 and to the Institute of Medicine in 1993. 

Professor Botstein has received numerous awards for his research excellence.  Notable among them are Eli Lilly and Company Award in Microbiology (1978), the Genetics Society of America Medal (1988), the Allan Award of the American Society of Human Genetics (1989), the Gruber Prize in Genetics (2003) and the Albany Medical Center Prize (2010).

Scott McIsaac is a 4^th year Ph.D. candidate in the Quantitative and Computational Biology Graduate Program at Princeton University. In 2010, Mr. McIsaac was awarded the prestigious National Science Foundation Graduate Research Fellowship. Mr. McIsaac received undergraduate degrees in physics and applied mathematics from Rice University in 2008.

 Sanford Silverman, Manager, Research Lab.  Sanford Silverman earned his Ph.D. in Molecular Biophysics and Biochemistry in 1979 from Yale University.  He is currently at the Lewis-Sigler Institute for Integrative Genomics in the Botstein lab.

Intellectual Property status

Patent protection is pending.

 Commercialization Strategy

Princeton University Office of Technology Licensing is pursuing a non-exclusive licensing strategy for the further development and commercialization of this technology.