NIH launches "Knockout Mouse Project"

Sep 7 2006

The National Institutes of Health (NIH) today awarded a set of cooperative agreements, totaling up to $52 million over five years, to launch the Knockout Mouse Project.

The goal of this program is to build a comprehensive and publicly available resource of knockout mutations in the mouse genome.  The knockout mice produced from this resource will be extremely useful for the study of human disease.

The NIH Knockout Mouse Project will work closely with other large-scale efforts to produce knockouts that are underway in Canada, called the North American Conditional Mouse Mutagenesis Project (NorCOMM), and in Europe, called the European Conditional Mouse Mutagenesis Program (EUCOMM). The objective of all these programs is to create a mutation in each of the approximately 20,000 protein-coding genes in the mouse genome.

"Knockout mice are powerful tools for exploring the function of genes and creating animal models of human disease. By enabling more researchers to study these knockouts, this trans-NIH initiative will accelerate our efforts to translate basic research findings into new strategies for improving human health," said NIH Director Elias A. Zerhouni, M.D. "It is exciting that so many components of NIH have joined together to support this project, and that the NIH Knockout Mouse Project will be working hand-in-hand with other international efforts. This is scientific teamwork at its best."

Knockout mice are lines of mice in which specific genes have been completely disrupted, or "knocked out." Systematic disruption of each of the 20,000 genes in the mouse genome will allow researchers to determine the role of each gene in normal physiology and development. Even more importantly, researchers will use knockout mice to develop better models of inherited human diseases such as cancer, heart disease, neurological disorders, diabetes and obesity. Recent advances in recombinant DNA technologies, as well as completion of the mouse genome sequence, now make this project feasible.

NIH today awarded five-year cooperative agreements totaling up to $47.2 million to two groups for the creation of the knockout mice lines. Recipients of those awards are

Regeneron Pharmaceuticals, Inc., in Tarrytown, N.Y., and a collaborative team from Children's Hospital Oakland Research Institute (CHORI) in Oakland, Calif.,the School of Veterinary Medicine, University of California, Davis (UC Davis); and the Wellcome Trust Sanger Institute in Hinxton, England.

In addition, NIH awarded another five-year cooperative agreement totaling $2.5 million to the Jackson Laboratory in Bar Harbor, Maine for the establishment of an NIH Knockout Mouse Project data coordination center. Finally, NIH awarded cooperative agreements to the University of Pennsylvania in Philadelphia and to the Samuel Lunenfeld Research Institute of Mount Sinai Hospital in Toronto to improve the efficiency of methods for creating knockout lines. Those agreements total about $2.5 million and run for three and two years, respectively.

"Building a genome-wide library of knockouts will require the skills of researchers from many different disciplines. We are confident that the multi-institution team we have pulled together will meet that challenge and deliver this much-needed resource into the hands of the worldwide research community," said James Battey, M.D., Ph.D., director of the National Institute on Deafness and Other Communication Disorders (NIDCD) and co-chair of the Trans-NIH Genomic Resources Working Group.

To date, academic researchers around the world have created mouse knockouts of about 4,000 genes. In addition, a random disruption strategy has been used by the International Gene Trap Consortium to mutate 8,000 mouse genes. Due to some overlap between these efforts, about 15,000 genes remain to be knocked out in the mouse genome.

The NIH program, along with NorCOMM and EUCOMM, intend to closely coordinate their efforts in order to avoid redundancy and maximize the efficiency of generating knockouts for all genes in the mouse genome.  Furthermore, the U.S., Canadian and European groups are committed to making their data and resources rapidly and openly available to researchers around the world.

"The international projects will exchange information and coordinate their efforts in much the same way that teams from many nations collaborated on the International Human Genome Project," said Colin Fletcher, Ph.D., a program director at the National Human Genome Research Institute (NHGRI), which will oversee administration of three of the five cooperative agreements that form the core of the Knockout Mouse Project.

Under its cooperative agreement, the team led by Pieter deJong, Ph.D., CHORI, along with K. C. Kent Lloyd, D.V.M., Ph.D., UC Davis; and Allan Bradley, Ph.D. FRS, and William Skarnes, Ph.D., at the Wellcome Trust Sanger Institute, plans to systematically create mouse embryonic stem (ES) cell lines in which 5,000 genes have been knocked out by gene targeting. The VelociGene division of Regeneron, led by David Valenzuela, Ph.D. and George D. Yancopoulos, M.D., Ph.D., will take aim at a different set of 3,500 genes. Both groups will utilize information from the finished mouse genome sequence to design targeting vectors, which will be built by large-scale, automated technologies. The combined collection of mouse ES cells with knockouts in 8,500 genes will be useful for producing knockout mice.

Other researchers will be able to obtain the ES cells and the vectors, which can be used to swiftly and efficiently to make live lines of knockout mice for use in biomedical studies. During the initial phase of the project, the ES cell lines and vectors used to mutate the genes will be available from the grantees who produced them. In addition, NIH is preparing to issue a solicitation for a program to implement a Knockout Mouse Project repository, which will be funded in the next year and through which all these materials will be available to the entire scientific community. 

Another crucial component of the effort will be the collection and coordination of data. Under the leadership of Martin Ringwald, Ph.D., the Jackson Laboratory will set up a Data Coordination Center for the Knockout Mouse Project. The center will collect information that will allow the research community to track the scheduling and progress of knockout production. The center will also serve as a central information resource for all publicly available knockout mutants and will integrate with other databases that contain mouse DNA sequence, additional information on mouse genetics and information on the physical and biochemical characteristics of the knockout mice.

Under two cooperative agreements administered by the National Institute on Drug Abuse (NIDA), Klaus Kaestner, Ph.D., and his colleagues at the University of Pennsylvania will focus on developing methods to create ES cell lines suitable for high-throughput gene targeting or trapping in C57BL/6, the strain of mouse used most widely by the scientific community. They will be joined in this effort by Andras Nagy, Ph.D., and his colleagues at the Samuel Lunenfeld Research Institute. In addition, Regeneron will receive funds to optimize its existing ES cell line for the C57BL/6 strain and its proprietary growth medium, both of which will be supplied to the CHORI-led team for use in the Knockout Mouse Project.

"Development of ES cell lines that can be used to make mutants in the C57BL/6 strain will be an important step forward in capitalizing on the vast amount of information obtained from years of research already done in this mouse strain," said NIDA Director Nora D. Volkow, M.D. 

While today's awards mark the official launch of the Knockout Mouse Project, NIH has been laying the foundation for several years. In the fall of 2003, NIH co-sponsored an international meeting that concluded that the time was right for a coordinated effort to produce knockouts in every mouse gene, and a commentary calling for such a project was published in the September 2004 issue of Nature Genetics, www.genome.gov/Pages/About/RecentArticles/AustinKnockoutMouseCommentary.pdf .

In October 2005, NIH and the U.K.'s Wellcome Trust took the first concrete step by awarding contracts that gave academic researchers access to a set of well-characterized knockout mouse lines created by Deltagen, Inc. of San Carlos, Calif., and Lexicon Genetics, Inc. of The Woodlands, Texas. NIH has expended about $11 million to acquire about 250 lines of these mice in the first year of the three-year contracts. Researchers can obtain information on what knockout mouse lines are available from this procurement and how to order them at: http://www.nih.gov/science/models/mouse/deltagenlexicon/list.html .

In June, NIH moved another step closer to its goal of a genome-wide library of knockout mice with the award of $800,000 to two public mouse repositories for the acquisition of existing knockout mouse lines that are not yet widely accessible to researchers. The award recipients were the Mutant Mouse Regional Resource Centers at UC Davis and the University of Missouri/Harlan in Columbia, both supported by the National Center for Research Resources (NCRR). NIH anticipates that more than 350 existing mouse lines will be deposited and made available to the research community over the next two years as a result of this effort. Researchers can obtain information on what knockout mouse lines are available from this effort and how to order them at: http://www.mmrrc.org/

The 19 NIH institutes, centers and offices contributing to the Knockout Mouse Project are: NCRR, National Eye Institute, NHGRI, National Heart, Lung and Blood Institute, National Institute on Aging, National Institute of Alcohol Abuse and Alcoholism, National Institute of Allergy and Infectious Diseases, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Child Health and Human Development, NIDCD, National Institute of Dental and Craniofacial Research, NIDA, National Institute of Environmental Health Sciences, National Institute of General Medical Sciences, National Institute of Mental Health, National Institute of Neurological Disorders and Stroke, National Institute of Diabetes and Digestive and Kidney Disease, National Cancer Institute, and the Office of AIDS Research.

For more information on the Knockout Mouse Project, go to www.nih.gov/science/models/mouse/knockout/ .  For a fact sheet describing what knockout mice are, how they are made and what they are used for, go to www.genome.gov/12514551 . To download a high-resolution photo of knockout mice, go to http://www.genome.gov/pressDisplay.cfm?photoID=5006 .

The National Institutes of Health – "The Nation's Medical Research Agency" – includes 27 institutes and centers, and is a component of the U.S. Department of Health and Human Services. It is the primary federal agency for conducting and supporting basic, clinical and translational medical research, and it investigates the causes, treatments, and cures for both common and rare diseases. For more, visit www.nih.gov .