Nucleic Acids for efficient conditional Inactivation and Mutagenesis of Eukaryotic Genes
Reference Number TO 02-00097
Challenge
A variety of methods for (un-)targeted mutagenesis and conditional inactivation of eukaryotic genes are known and well established. However, all state-of-the art technologies suffer from a set of disadvantages including time – and cost consuming cloning procedures, generation of defective translation products with unpredictable effects, required knowledge of intronphases and open reading frames etc. and there is an actual demand for new and refined technologies to accelerate research and development.
Technology
By making use of „nonsense mediated mRNA decay (NMD)“,the technology discloses nucleic acid constructs for (multiple) conditional inactivation/reactivation of genes enabling efficient targeted mutagenesis or for nondirectional mutagenesis (eg. gene traps). The technology particularly relates to targeted mutagenesis of genes in non-human ES cells and non-human mammals producible therefrom. NMD is a mechanism that controls the gene expression in eukaryotic cells by early degradation of incorrect mRNA molecules with untimely stop codons. The inventive constructs mediate NMD and by integration of stop codons in all three open reading frames allows for inactivation of genes comprising at least one intron. The NMD mediating exon is flanked by intelligent functional intron sequences allowing for inversion or deletion of dedicated sequence sections and thereby enabling conditional regulation of the target gene. The key advantage of this technology is the time and effort reduction that can be achieved when constructing reliable transgenic vectors and animal models.
Commercial Opportunity
The technology is available for in-licensing for further development in transgenic applications.
Developmental Status
Various nucleic acid constructs were developed for use in eukaryotic cells and mice.
Patent situation
A priority application was filed in June 2007, PCT/EP application is pending. The application is published under WO 2008/155306 A1.
Further Reading
Authorized thesis Dr. Nawrath: www.digibib.tu-bs.de
