Targeted Mutagenesis of the Zebrafish Genome Through Manipulation of Helicase Activity

Description (Set) Helicase enzymes are important enzymes functioning in DNA replication, repair, recombination, and RNA transcription. RecQ helicases are a family of helicases that act to suppress homologous recombination at specific junctions. Despite the many advantages of the zebrafish as an experiment organism, there is currently no method available to induce targeted mutations in the zebrafish genome. Scientists at Johns Hopkins University are developing methods to perform targeted mutations in the germline of zebrafish embryos through the manipulation of the activity of the RECQL family of DNA helicases, allowing for the analysis of gene function and the creation of models for human genetic diseases. Mutants which are created in this manner can be used to examine the pathophysiology underlying specific diseases, to test potential drug therapies, or as the substrate for large-scale screens of small molecules to identify new therapies. Importantly, this invention may also be adapted for equivalent use in other vertebrate or invertebrate model organisms. Proposed Use (Set) Current available technologies allow for targeted mutations in mice through gene knockouts, knockins, and chromosomal engineering. This same technology is not currently available for zebrafish. Also in zebrafish, there is no genetic method to generate mosaic animals (that contain only small clones, or groups, of mutant cells), or to easily generate embryos lacking the maternal product of a gene essential for normal development. Through manipulation of the activity of the RECQL family of DNA helicases, either in the entire embryo or specifically in germ cells, we can induce high rates of mitotic recombination, thus facilitating all of the above approaches in zebrafish.

Inventor(s): Fisher, Shannon ,Inventor (Set) Fisher, Shannon ,McCallion, Andrew Smythe,Xie, Jing

Type of Offer: Licensing

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