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NBRP Fundamental Technologies Upgrading Program Project's Progress


Focus Production of conditionally gene-disrupted ES-cell clones and establishment of a
database for the inactivated genes
Principal Investigator Yasumasa Ishida   Graduate School of Biological Sciences Nara Institute of Science and Technology
Project Period FY2010–2011
Overview   In order to disrupt all the protein-coding genes in the mouse embryonic stem (ES) cell by using the insertional-mutagenesis techniques and to freely distribute a tremendous number of ES-cell mutants generated for researchers in the academic fields, the knockout mouse project (KOMP) has been carried out in some of the major western countries. However, random and conditional disruption of transcriptionally silent genes in ES cells through gene trapping has been poorly developed in the project based on the conventional gene-trapping technologies. Thus, people have already started accelerating to disrupt a large number of the remaining ‘difficult-to-trap’ genes individually by using gene-targeting (non-random) strategies.

  In this program, we will try to attain quick, selective, and conditional trapping of transcriptionally silent genes in mouse ES cells by establishing a novel strategy in which the diphtheria toxin (DT)-mediated negative selection is involved and by exploiting the UPATrap methodology that had already been refined thoroughly in the national bio-resource project (NBRP) of Japan during its fiscal years H19-H20. Further, we will also establish a fully competent and useful database for genes disrupted in an enormous number of ES-cell clones generated in the current program. By combining these novel developments effectively, we would be able to create a highly useful resource for a wide range of biomedical researchers, and consequently, Japan would be able to occupy a unique position in the international collaborative effort of the KOMP.
Project's Progress