Transgenic technologies are prohibitively expensive for individual laboratories and require extremely skilledand dedicated personnel and institutional support. This is because first, many factors greatly affectexpression levels of randomly inserted DNA, which leads to transgenes being transcriptionally silent at manychromosomal locations. In addition, there is a considerable risk as to whether targeted ES cells willcontribute to the germline. As a result, no transgenic facility will guarantee transgene expression, nor thattargeted ES cells will go to germline. Secondly, many investigators whose studies require the use oftransgenic animals are not trained in their generation. This is partly the case for this program project wherethe expertise of investigators and their lab personnel range from extensive experience to very littleexperience. Therefore, the expertise that is available in the transgenic core at UMKC has also beenincluded in this program project. The Director and Co-Director will assist the principal investigators in thedesign of constructs, educate personnel and students, and oversee the general operation of the core. Theservices of this core will not only be to generate transgenic mice, but to cross breed mice to generate thecorrect genotype, and to maintain the mice until ready for the Mechanical Strain Core or to be sent to theinvestigator. The purpose of this core is to provide efficient and economical service to the members of theProgram Project plus the needed educational component. The core will also provide the following: 1)generation of transgenic embryos for screening of expression of Dmp1 and Mepe promoter fragments forosteocyte selectivity and response to mechanical load; 2) generation of transgenic mice expressing the 8and/or 10 kb Dmp1-Cre, intact MEPE or MEPE promoter fragments, 3). generation of wild type Cx43 andtruncated Cx43 mice and their targeted expression in osteocytes; 4) generation of Dmp1-loxP and E-11-loxPmice; 5) generation of floxed mice lacking Dmp1 or E11 in osteocytes using various promoters regulatingCre expression; and 6) maintainance of all Cre mice needed for this project, including osteocalcin-Cre, 8(and/or 10) kb Dmp1 Cre, and 3.2 kb Col 1a1 ERt2-Cre mice. In addition, the Core will serve as a resourcefor consultation and training for prinicipal investigators, their students and personnel for genotyping andscreening of transgenic and knockout mice.
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