Overall Component This project supports continued funding for the National Xenopus Resource (NXR) located at the Marine Biological Laboratory in Woods Hole, MA and identified as one of two top priorities for the Xenopus community. The NXR was established in 2010 to serve as a national resource for researchers working with the Xenopus amphibian model system, which includes two species Xenopus laevis and Xenopus tropicalis. The NXR serves as a national repository for community-generated animal stocks as well as serving as an advanced training venue that greatly facilitates productivity for a multitude of researchers. It is a community-oriented resource center that serves all Xenopus researchers, including individuals from large research-focused universities to small liberal arts colleges. As biological research becomes ever more complex, often requiring specialized animal lines and involving diverse technologies, individual laboratory units become insufficient to meet all of the demands required for addressing significant biological problems and a centralized repository or stock center becomes essential. There are five main aims to this grant. First, the NXR will maintain current stocks of frogs as well as obtain new lines making them available to the community; this includes special inbred, wild type and mutant lines of both species. In the second aim we outline the NXR custom research services for creating transgenic and mutant animals that are offered to the research community. In the third aim we outline the resources that we have created for the Xenopus community that helps to facilitate the use of new techniques in the community. These resources benefit from the excellent MBL facilities for research, teaching and housing. This includes the development of new advanced training workshops taught by experts in each field that serve to teach and propagate specialized techniques; this includes workshops on bioinformatics, imaging, genome editing, transgenesis and husbandry. In this aim we also outline the research hotel service, which provides opportunities for researchers to come to the MBL for short-term visits and use NXR resources. The last two aims are focused on applied research.
In Aim 4 we describe new transgenic technology and enhancement of the new genome editing technology. In the last aim we propose to optimize sperm cryopreservation in X. laevis and develop a new health screening PCR service to test for the presence of specific pathogens.
Research using the amphibian Xenopus, because of unique advantages as an experimental system, has revealed key insights in many domains of biomedical research, including cell biology, development, neurobiology, physiology and signal transduction, achievements that are supported by having a centralized National Xenopus Resource for raising and distributing animals and disseminating the most current technology to the research community. In each of these areas research has led to significant insights about the causes of human diseases, including cancer, birth defects, diabetes and neurological disease, and provided key underpinnings for the field of regenerative medicine.
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|DeLay, Bridget D; Corkins, Mark E; Hanania, Hannah L et al. (2018) Tissue-Specific Gene Inactivation in Xenopus laevis: Knockout of lhx1 in the Kidney with CRISPR/Cas9. Genetics 208:673-686|
|Pearl, Esther; Morrow, Sean; Noble, Anna et al. (2017) An optimized method for cryogenic storage of Xenopus sperm to maximise the effectiveness of research using genetically altered frogs. Theriogenology 92:149-155|
|Ratzan, Wil; Falco, Rosalia; Salanga, Cristy et al. (2017) Generation of a Xenopus laevis F1 albino J strain by genome editing and oocyte host-transfer. Dev Biol 426:188-193|
|Savova, Virginia; Pearl, Esther J; Boke, Elvan et al. (2017) Transcriptomic insights into genetic diversity of protein-coding genes in X. laevis. Dev Biol 424:181-188|
|Webb, Bryn D; Metikala, Sanjeeva; Wheeler, Patricia G et al. (2017) Heterozygous Pathogenic Variant in DACT1 Causes an Autosomal-Dominant Syndrome with Features Overlapping Townes-Brocks Syndrome. Hum Mutat 38:373-377|
|Tandon, Panna; Conlon, Frank; Furlow, J David et al. (2017) Expanding the genetic toolkit in Xenopus: Approaches and opportunities for human disease modeling. Dev Biol 426:325-335|
|Wlizla, Marcin; Falco, Rosalia; Peshkin, Leonid et al. (2017) Luteinizing Hormone is an effective replacement for hCG to induce ovulation in Xenopus. Dev Biol 426:442-448|
|Sedzinski, Jakub; Hannezo, Edouard; Tu, Fan et al. (2016) Emergence of an Apical Epithelial Cell Surface In Vivo. Dev Cell 36:24-35|
|Vukovi?, Lidija D; Jevti?, Predrag; Zhang, Zhaojie et al. (2016) Nuclear size is sensitive to NTF2 protein levels in a manner dependent on Ran binding. J Cell Sci 129:1115-27|
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