Objective
The ultimate goal of this project is to develop molecular methods leading to integration and expression of new beneficial transgenes in fish and biological containment of these fish, with the ability to transmit these trait under controlled conditions. This goal will be achieved through three distinct objectives to the work plan in this proposal. The first and primary objective of the project is the production and of analysis of stable is of lines of fish which have been rendered transgenically sterile by the inhibition of gonadotropin releasing hormone (GnRH) at the level of the brain. This objective will be achieved by the expression of GnRH antisense and/or ribozyme RNAs ("anti-messages") to inhibit the biosynthesis of GnRH. Absence of GnR will result in a blockage of the hypothalamo-pituitary-gonad axis. The second objective is to assess the effectiveness of the induced sterility. This will be achieved by analysis of the reproductive endocrine system and the functionality of the gonads. Furthermore, in order to produce stable lines of transgenic fish, methods to regenerate gametogenesis in these sterile transgenic fish (a key element required to produce fish lineages) will be developed. The third objective is to produce transgenic fish expressing new potentially beneficial genetic traits (hyperprolactinemic), in addition to biological containment. It is proposed to use three species of fish in this project; zebrafish (DaMiorerio), rainbow trout (Oncorhynchus mykiss) and tilapia (Oreochromis niloticus).
The major objectives in this proposal will be achieved through 4 interrelated and interdependent tasks. The first task will involve optimization of stable integration and expression of transgenes. To this end, strong all tissue and tissue specific promoters, enhancer sequences, NLS peptides and 3'UTR stabilisers will b isolated. In the second task these sequences will be combined into constructs with the GnRH anti-message and also reporter genes, such as GFP and lac Z. Furthermore, to assess the efficiency of methods used t shut off GnRH expression in-vitro, GnRH expressing cell lines will be established. The third task will involve the production and characterization of the transgenic sterile fish. The GnRH content will be measured in discrete brain areas and pituitary using a specific and sensitive ELISA technique. The functionality of the gonads will be studied in trout and tilapia by histology and by measuring steroid blood plasma levels by specific RIA. The fourth task will involve the combination of a potential beneficial genetic trail (hyperprolactinemia) with biological containment. Finally, we plan to conduct an evaluation of gene flow within salmonid and tilapia species. Attempted breeding will be used to determine if gene flow can OCCUI between male transgenic stelile fish and females from the same or related species.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences biochemistry biomolecules
- natural sciences biological sciences genetics RNA
- medical and health sciences clinical medicine endocrinology
- natural sciences biological sciences histology
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Coordinator
90 GALWAY
Ireland
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