Li Y et al. (1997) International C. elegans Meeting "CONTINUING IMPROVEMENTS IN TECHNIQUES FOR PREPARATION OF C. ELEGANS FOR TRANSMISSION ELECTRON MICROSCOPY"

Li Y, Kimble JE

[International C. elegans Meeting, 1997]

We have been continuing to work on protocols for fixation, staining and embedding and want to share our results with the nematode community. Representative micrographs will be shown and protocols will be available for anyone interested.

Yeung MH et al. (1997) International C. elegans Meeting "THE USE OF C. ELEGANS FOR INSECTICIDE DISCOVERY"

Hodgkin JA, Yeung MH, Robinson MP

[International C. elegans Meeting, 1997]

C. elegans can be used as a tool in the discovery of new insecticides. Lead Detection - Wild type C. elegans can be used for screening of unknown compounds. The ideal is to use the target species on its normal host plant. However, that may not be amenable to screening. C. elegans has its advantages: - short life cycle and ease of culturing means the C. elegans can be grown to a large number in a short time using inexpensive culture medium. It can be stored frozen between use. - Our initial study shows that C. elegans has a reasonably selective activity to a spectrum of insecticide standards. Mode Of Action (i) - a panel of C. elegans mutants (each with known resistance or sensitivity) can be used to define the mode of action of unknown compounds. - If a mutant responds in a characteristic way to a compound, it is indicative of a relevant mode of action. - If all mutants do not provide any characteristic response to a compound, it is indicative of a novel mode of action. - Behavioural symptoms can be very useful. - It can be useful for predicting resistance and cross-resistance. - It can be useful for grouping unknown compounds. Mode Of Action (ii) - the unknown compound with a putative novel mode of action can be used as selective agent after EMS mutagenesis to generate mutants. - It can be useful towards understanding the mode of action by carrying out further work on the mutant. - The existing extensive information on ACeDB will facilitate characterisation of the mutant. - The mutant can be screened against a spectrum of putative relevant mode of action insecticide standards to check for cross resistance.

Fire A et al. (1997) International C. elegans Meeting "SOME PROMOTER ACTIVITY PATTERNS"

Hsieh J, Macarah C, Fire A, Kelly B, Harfe BD, Montgomery MK, Xu SQ

[International C. elegans Meeting, 1997]

We will describe a variety of interesting germline and somatic promoter activity patterns and make some suggestions as to how you might be able to engineer your gene to be expressed in any of all of these patterns.

Hughes D et al. (1998) European Worm Meeting "Systematic gene inactivation in C. elegans"

Hughes D, Jones SJ, Durbin RM, Coulson AR

[European Worm Meeting, 1998]

We have initiated a project to investigate the applicability of chemical mutagenesis to large-scale targeted gene inactivation in C.elegans (see abstract by Plasterk et al). In collaboration with the Plasterk lab and co-operating with a number of other like-minded labs we are aiming to develop an efficient process whereby publicly available knock-outs of all genes would be the objective. It is anticipated that, initially at least, targeted genes will be largely request driven. Current methodology will be presented.

Melissa J Kirven-Brooks et al. (2003) International Worm Meeting "A C. elegans Experiment on the International Space Station"

Kristina Lagel, Jeanne Hale, Kevin Sato, Melissa J Kirven-Brooks, Delia Santiago, June Sunga, Debra Reiss Bubenheim

[International Worm Meeting, 2003]

An Incubator will be available on the International Space Station with capabilities that are well suited to examine the effects of microgravity and cosmic radiation on C. elegans. The Space Station Biological Research Project (SSBRP) Incubator provides temperature control between 4 and 45 degrees C, and has commandable data, power and video ports to support life science experiments. For the first Incubator flight (slated for launch in 2005) an experiment is planned with C. elegans, to examine growth, development, reproduction and behavior, as compared to ground-based controls. The C. elegans will be incubated in liquid axenic medium 1 in OptiCell (trademark) containers. During the 90 day increment, cultures will be subcultured and videotaped regularly, and live, chemically preserved and frozen samples will be collected. These samples will be analyzed for changes at the genetic and protein level. Ground studies will be presented on baseline growth and behavior of C. elegans in this hardware. Reference: 1 Lu, NC; Goetsch, KM. Carbohydrate requirement of Caenorhabditis elegans and the final development of a chemically defined medium. Nematologica 39(3):303-311, 1993

J T Stenvall et al. (2002) European Worm Meeting "Characterisation of two EF-1a homologues: translational elongation factors SelB and cgp-1."

J T Stenvall, Simon Tuck, O P Persson

[European Worm Meeting, 2002]

Selenoproteins are usually involved in redox reactions. Most eukaryotic selenoproteins are believed to be necessary for the protection of the cell against oxygen radicals.

Jui-ching Wu et al. (2002) West Coast Worm Meeting "Genetic Analysis of Asymmetric LET-99 Localization"

Meng-Fu Bryan Tsou, Lesilee S Rose, Jui-ching Wu

[West Coast Worm Meeting, 2002]

Asymmetric cell division, which generates two daughter cells with different cell fates, is important for normal development. Two events are necessary for cells to divide asymmetrically. First, polarity has to be established to position cell fate determinants asymmetrically. Second, the mitotic spindle has to be properly oriented to correctly segregate cell fate determinants into different daughter cells.

Vidal MI (1997) International C. elegans Meeting "EARLY STEPS IN THE CONSTRUCTION OF A PROTEIN-PROTEIN INTERACTION DATABASE FOR C. ELEGANS"

Vidal MI

[International C. elegans Meeting, 1997]

Large-scale projects of genome and Expressed Sequence Tags (ESTs) sequencing are extremely valuable. One of the next challenges is to determine how the predicted ORFs resulting from such projects can be functionally linked to each other. Since protein-protein interactions are critical to such a wide range of processes, I propose to functionally link ORFs by identifying interaction partners and relate these interactions to biological questions. Automated high-throughput strategies will be presented to generate worm "Interaction Sequence Tags" (ISTs). ISTs are defined as pairs of short sequences obtained from cDNAs encoding interacting proteins in the yeast two-hybrid system. ISTs will be released in the form of databases publicly available on the Internet. In addition, automated high-throughput strategies will also be presented to generate !interaction defective alleles!. These alleles are defined as single amino-acid changes that specifically affect protein-protein interaction and can be selected in the reverse two-hybrid system (Vidal et al, 1996, PNAS, 93, 10315-10320 and 10321-10326). Such interaction defective alleles will be made available to the worm community. They could be tested for their ability to rescue particular phenotypes when reintroduced in the relevant worm loss-of-function mutants. This work should help developing the tools and the strategies needed to eventually generate and interpret a comprehensive protein-protein interaction map based on both physical and genetic interactions.

Memar, Nadin et al. (2009) International Worm Meeting "A screen for temperature-sensitive embryonic lethal mutations affecting cell focussing in Caenorhabditis elegans."

Wiekenberg, Anne, Schnabel, Ralf, Martin, Katharina, Memar, Nadin

[International Worm Meeting, 2009]

Schnabel et al. (2006) proposed cell focussing as mechanism for global cell sorting by cell identity in the C. elegans embryo. It should be helpful to find mutants to define the mechanism of this process. Interestingly a very large collection of nonconditional embryonic lethal mutants did not contain a mutant affecting cell focussing. We supposed that genes affecting this process might be highly pleiotropic, since it should also keep larvae and adults in shape and/or maternal rescue may occur in the embryo. Therefore, ts mutants may be the tool of choice to identify genes. We designed a new screen for ts mutants using the worm sorter, since a very large number of mutants may be required but the yield is less than 1%. Identification of specific mutants is very cumbersome since candidates have to be subjected to a careful 4D analysis. Only mutants in which fates are wild-type - but the positioning of subsets of cells is wrong - qualify as potential cell focussing mutants. A first screen yielded two candidates in approx. 1000 ts mutants. In the next round we isolated again 1000 mutants, which now await analysis. We expect that up 10000 ts mutants may be required to define a cell focussing pathway. However, the mutant collection may be also a valuable resource for the worm community.

Hu, Yan et al. (2013) International Worm Meeting "Treat worm infections with crystal protein expressing in probiotic like bacteria."

Ellis, Brian, Huerta, Daniel, Miller, Melanie, Pogliano, Joseph, Aroian, Raffi, Derman, Alan, Hu, Yan

[International Worm Meeting, 2013]

Soil-transmitted helminths (namely hookworms, whipworms, and Ascaris large roundworms) are intestinal nematodes, which cause diseases of poverty that infect upwards of two billion people worldwide. These parasites are a major threat to health and development of hundreds of millions of children and pregnant women. Enormous hurdles must be overcome in order to develop and deliver urgently needed new therapies (anthelmintics) to replace old ones that perform sub-optimally and are losing efficacy. Any new therapy must be extremely cheap, be able to be produced in tremendous quantities to treat hundreds of millions of people, have a stable shelf life, and be capable of storage and delivery under adverse environmental conditions. Our research has uncovered a radical and unique new approach that solves each of these challenges: expression of vertebrate-safe, anthelmintic (anti-nematode) proteins in "probiotic-like" food-grade bacteria. Such bacteria can be produced cheaply, in great quantity, stored stably, and delivered under adverse conditions. Here we will discuss our work to develop such engineered bacterial therapy using the anthelmintic crystal protein Cry5B normally made by Bacillus thuringiensis (Bt). We will present data on how Cry5B can be expressed in a non-Bt bacterium related to food-grade bacteria, and the strong efficacy of such a bacterium in clearing hookworm infections in rodents. We will also update progress on engineering several food-grade bacteria to express Cry5B as a critical step towards implementation of this novel anthelmintic approach.