The Expression Core will determine the developmental times and tissues in which candidate genes identified by Projects I, II and III are expressed. These studies will determine both RNA and/or protein expression patterns using immunohistochemistry or in situ hybridiation, whichever is deemed appropriate. Further, genes of particular interest will undergo analysis using the RCAS-TVA system for tissue expression of both wildtype and dominant negative proteins. Diaphragm whole mounts are difficult to examine, but the director and staff of this core have learned the necessary techniques from Dr. Akerman. Lung organ culture has been in use in the lab for 20 years. RNAi technology has been used by the Pis for the past 8 years. Together with the Drosophila Core, RNAi technology will be uniquely available for the program project. Brief descriptions of the protocols that Dr. Loscertales uses routinely which will be instrumental in analyzing CDH candidate genes follow.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
5P01HD068250-04
Application #
8708178
Study Section
Special Emphasis Panel (ZHD1-DSR-N)
Project Start
Project End
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
4
Fiscal Year
2014
Total Cost
$90,832
Indirect Cost
$50,846
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Longoni, Mauro; High, Frances A; Russell, Meaghan K et al. (2014) Molecular pathogenesis of congenital diaphragmatic hernia revealed by exome sequencing, developmental data, and bioinformatics. Proc Natl Acad Sci U S A 111:12450-5
Lage, Kasper (2014) Protein-protein interactions and genetic diseases: The interactome. Biochim Biophys Acta 1842:1971-1980
Lundby, Alicia; Rossin, Elizabeth J; Steffensen, Annette B et al. (2014) Annotation of loci from genome-wide association studies using tissue-specific quantitative interaction proteomics. Nat Methods 11:868-74