The DTCC Consortium: Year One Update
Feb 07 2013
The DTCC consortium is a joint research effort by the University of California, Davis (UC Davis), Toronto Center for Phenogenomics (TCP), Children’s Hospital Oakland Research Institute (CHORI) and Charles River. It is funded by the National Institutes of Health (NIH) as part of the second phase of the Knockout Mouse Project (KOMP2) to create a repository of knockout (KO) mouse lines and phenotype data. This repository will provide researchers with a valuable resource to develop better models of human diseases such as cancer, heart disease, neurological disorders, diabetes and obesity. The target genes for production and phenotyping were carefully chosen, with primary emphasis on genes with little to no annotation (i.e., 0-2 GO terms, etc.) and genes in which investigators had registered interest. All models created will have a battery of phenotyping assays performed on them to generate data that can be useful in determining a particular model’s suitability for a specific research application.
DTCC models are created at UC Davis and TCP via embryonic stem cell injection or, increasingly, via a highly efficient morula aggregation technique. Once Cre excision has been completed to produce the lacZ-tagged KO mice, Charles River receives the lines for in vitro fertilization (IVF) to expand the line, standardize the health status of all lines and assess each line for viability. After IVF, 7 homozygous males, 7 homozygous females and a wild-type male and female from adult viable lines are delivered to a renovated, dedicated facility at UC Davis, where all equipment for assays in the adult pipeline has been installed and is now available. LacZ analysis for gene expression and all adult pipeline phenotyping is performed at this facility. All subviable lines are delivered from Charles River to TCP as heterozygous animals for additional breeding to generate embryos or neonates that proceed through an image- and pathology-based pipeline to screen for phenotype.
The DTCC has accomplished a great deal during this first year, including ~500 microinjections and identification of > 100 germline-positive KO mouse lines. To date, ~60 KO mouse lines have been sent to Charles River for expansion, and cohorts for 5 KO lines have entered the comprehensive phenotyping platform.
The DTCC has also been proactive in driving technology development to further optimize its processes as well as ensure the delivery of its commitments to the KOMP2 project. Scientists at TCP have developed protocols for the efficient use of morula aggregation and recently offered a workshop in which members of the International Mouse Phenotyping Consortium (IMPC) were trained in the new methodology. In addition, TCP has tested Cre excision lines to enhance the efficiency of Cre excision by maternal effect excision and has developed an in vitro method for Cre excision in mouse embryos.
To explore the repository’s potential research applications, the DTCC will be sharing its progress at upcoming scientific meetings, including round table discussion panels at Experimental Biology in April 2013 and American Society for Human Genetics in October 2013. Consortium members will also present at educational seminars, including the 2013 Charles River Short Courses in the US and Europe.
To learn more about the DTCC’s recent work, please review the multimedia resources below. If you have additional questions, we invite you to email dtcc@komp.org. For questions regarding Charles River involvement in the DTCC, please email us at askcharlesriver@crl.com.
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