Scientists are limited with the kinds of experiments that can be done with humans. Therefore, to understand human biology, investigators must study related animals. Remarkably, studies of even distantly related animals like soil round worms can provide valuable information about human biology. One such soil round worm, the nematode Caenorhabditis elegans (C. elegans), is so useful experimentally and so closely related to humans, that it is one of the most intensively studied animals in biological research.
One very important method used to study the nematode is genetics. Genetics involves the study of mutants or animals with abnormal genes that cause them to be different anatomically, physiologically, or developmentally from normal animals. Unlike with humans, scientists can treat nematodes with chemicals that cause mutations. By using the tricks of the geneticist, the genes that are affected in the mutant can then be identified. Finally, by studying what goes wrong when a gene is defective, the normal function of the gene can be understood.
Genes are made of deoxyribonucleic acid (DNA), a simple string of four different chemicals, that when read by the cellular machinery serves as a template to make proteins and other parts of the cell. Scientists have discovered the sequence of the DNA in many different organisms, including the nematode C. elegans and humans. From the DNA sequence, all the genes in an organism can be determined. Also, when a gene sequence in the nematode is similar to that in humans, this means that the genes are probably related, and that studies of the nematode gene will tell us something about the function of the human gene.
The sequence of the entire string of DNA in the nematode, which is known collectively as its genome, reveals that this animal has a total of 20,000 genes. In contrast, humans have about 35,000 genes. Of the 20,000 genes in the nematode, 2,000 have been studied using genetic methods. There is very little known about the function of the remaining 18,000 genes because mutant versions have not yet been isolated that can be used in genetic studies. The major research efforts of my laboratory are directed at producing mutant versions of these 18,000 genes.
Our goal is to construct 18,000 mutant nematode strains, each of which is mutant for a single gene. We use chemical and ultraviolet light treatment to cause mutations in the DNA of the nematodes. To identify the mutations, we use a method called the polymerase chain reaction (PCR), the same method used to fingerprint the genomes of suspects in a criminal investigation. Animals with mutations in a single gene are rare, and to find them requires many thousands of PCR tests. At present, we do 50,000 PCR tests each week to isolate about 20 mutant strains. To perform this number of tests, we use a combination of highly skilled staff, robotics, and other instrumentation.
To date, we have produced nearly 2,000 mutant nematode strains. Every strain that is produced is made available for further study to researchers around the world. Our mutant strains are now being used to study a wide range of biomedical problems, from cancer to neurological disease.
Education
B.A., University of Colorado at Boulder, 1979
Ph.D., Northwestern University, Evanston, IL, 1983
Honors and Awards
1975 University of Colorado Regents Scholarship
1983-1985 Muscular Dystrophy Association Fellowship
Memberships
American Society for Cell Biology
Joined OMRF Scientific Staff in 1993.
Since the completion of the genome sequence, it has become clear that traditional forward genetic methods are not sufficient for a comprehensive understanding of all 20,000 of these genes. In response to the limitations of forward genetic methods, my laboratory developed technologies for targeted gene inactivation. Our methods are used worldwide in many individual labs. Although one might leave it up to these individual labs to knockout the function of each of the 20,000 C. elegans genes, four years ago we proposed to the NIH that a systematic, genome-wide gene knockout project, performed by a few labs able to devote the time and resources to the project, would greatly accelerate C. elegans biology and the many biomedical research projects for which C. elegans could provide a model system. This proposal was accepted, and four years ago we formed the C. elegans Gene Knockout Consortium.
It is already clear that our gene knockout resource is immensely valuable to the C. elegans research community. Each strain that we make is deposited in C. elegans Genetics Center where they are accessible to any investigator. Over half of the new strains submitted to the CGC in the last year were from our lab. Further, the CGC filled over 2,000 requests for these strains, accounting for nearly 20 percent of the total strains shipped last year by the CGC. Further, we have numerous examples showing that the resource is especially valuable to those colleagues studying other organisms who wish to exploit C. elegans. Our knockout resource provides a rapid and convenient entry into a model system that allows investigators to bring to bear many unique genetic methods to address significant issues in biomedical and clinical research. By encouraging others to enter the field, especially others who bring the field new experiences and expertise, we accelerate the pace of discovery of protein function in C. elegans and in vertebrates.
Finally, our work allows clinical researchers to exploit C. elegans to study the basic cell biology of several human diseases including polycystic kidney disease, cancer and muscular dystrophy.
Two small molecule drug-like compounds that reduce the function of the mitotic spindle checkpoint in human cells have been shown to have effects on the nematode Caenorhabditis elegans. A better understanding of these compounds may lead to the development of a new class of anti-cancer, chemotherapeutic agents. We are collaborating with Dr. Gary Gorbsky to discover the mechanism of action of these compounds by 1) examining their phenotypic effects on the nematode and 2) through genetic screens to discover putative target genes/pathways that when mutant can render animals resistant to their affects. Using available antibodies and GFP-tagged proteins, first we are characterizing the arrest phenotype induced by these compounds in wild type C. elegans and in animals with mutations in genes with known roles in mitosis or the cell cycle. Second, we have already identified a mutant with a mild chromosome non-disjunction phenotype that is resistant to one of the two compounds described here. We are screening for others which will be mapped genetically and physically to identify the responsible genes and their encoded products. Finally, we have discovered a paradoxical interaction between one of these compounds and the well studied canonical programmed cell death pathway in C. elegans. We are studying the basis for this fascinating interaction to discover the relationship between compound activity and programmed cell death. Understanding this relationship may provide entrées to the development of new therapies that either suppress or activate apoptotic pathways so as to either protect cells from damage or eliminate cells that are diseased.
Recent Publications
Moulder GL, Cremona GH, Duerr J, Stirman JN, Fields SD, Martin W, Qadota H, Benian GM, Lu H, Barstead RJ. alpha-actinin is required for proper assembly of Z-disk / focal-adhesion-like structures and for efficient locomotion in Caenorhabditis elegans. J Mol Biol 403:516-528, 2010. [Abstract]
Flibotte S, Edgley ML, Chaudhry I, Taylor J, Neil SE, Rogula A, Zapf R, Hirst M, Butterfield Y, Jones SJ, Marra MA, Barstead RJ, Moerman DG. Whole-genome profiling of mutagenesis in Caenorhabditis elegans. Genetics 185:431-441, 2010. [Abstract]
Moerman DG, Barstead RJ. Towards a mutation in every gene in Caenorhabditis elegans. Brief Funct Genomic Proteomic 7:195-204, 2008. [Abstract]
Selected Publications
Crittenden SL, Bernstein DS, Bachorik JL, Thompson BE, Gallegos M, Petcherski AG, Moulder G, Barstead R, Wickens M, Kimble J. A conserved RNA-binding protein controls germline stem cells in Caenorhabditis elegans. Nature 417:660-663, 2002. [Abstract]
Edgley M, D’Souza A, Moulder G, McKay S, Shen B, Gilchrist E, Moerman D, Barstead R. Improved detection of small deletions in complex pools of DNA. Nucleic Acids Res 30:e52, 2002. [Abstract]
Tobin D, Madsen D, Kahn-Kirby A, Peckol E, Moulder G, Barstead R, Maricq A, Bargmann C. Combinatorial expression of TRPV channel proteins defines their sensory functions and subcellular localization in C. elegans neurons. Neuron 35:307-318, 2002. [Abstract]
Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O’Rourke J, Mole DR, Mukherji M, Metzen E, Wilson MI, Dhanda A, Tian YM, Masson N, Hamilton DL, Jaakkola P, Barstead R, Hodgkin J, Maxwell PH, Pugh CW, Schofield CJ, Ratcliffe PJ. C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl Hydroxylation. Cell 107:43-54, 2001. [Abstract]
Hao JC, Yu TW, Fujisawa K, Culotti JG, Gengyo-Ando K, Mitani S, Moulder G, Barstead R, Tessier-Lavigne M, Bargmann CI. C. elegans slit acts in midline, dorsal-ventral, and anterior-posterior guidance via the SAX-3/Robo receptor. Neuron 32:25-38, 2001. [Abstract]
Genetic Models of Disease Research Program, MS 48
Oklahoma Medical Research Foundation
825 N.E. 13th Street
Oklahoma City, OK 73104
Phone: (405) 271-1766
Fax: (405) 271-7312
E-mail: barsteadr@omrf.org
John Rummage, Ph.D.
Manager Laboratory
Gene Chao
Network Support Specialist
Gerry McElroy
Laboratory Assistant
