KOMP Phenotyping Data

The initial data are coming from a Recovery Act (ARRA) supported project to produce and characterize 312 mutant knockout lines produced primarily from KOMP targeted ES cells. The phenotype screen in adult mice includes LacZ reporter gene tissue distribution in all mutant lines, as well as fertility and viability, necropsy, behavioral phenotypes and a comprehensive analysis of transcriptome changes in homozygous mutant mice for 100 of the mutant lines. View the current KOMP Phenotyping gene list

• Whole-mount Lacz Annotations

  Adult mice are used to map the expression profile of genetically inserted reporter gene (lacZ) following the theory and protocol laid out in Adams and Gale (Mammalian & Avian Transgenesis, S. Pease & C. Lois, Eds., Springer-Verlag, 2006). Mice are fixed by cardiac perfusion using a saline rinse followed by 4% paraformaldehyde (PFA). After the tissues are dissected and rinsed in phosphate buffered saline (PBS), the tissues are post-fixed in 4% PFA for 30 minutes. Tissues are then rinsed in PBS and stained in lacZ staining solution for 48 hours. Following staining, tissues are post-fixed in neutral buffered formalin (NBF), cleared overnight in 50% glycerol, and then undergo a final clearing and are stored in 70% glycerol with 0.01% sodium azide. Tissues are placed in fresh 70% glycerol and examined using the Leica M165C dissecting microscope. All lacZ staining is photodocumented using a Leica DFC425 digital camera and noted as part of the annotation information.

• Frozen Lacz Annotations

  Adult mice are used to map the expression profile of genetically inserted reporter gene (lacZ) following the theory and protocol laid out in Adams and Gale (Mammalian & Avian Transgenesis, S. Pease & C. Lois, Eds., Springer-Verlag, 2006). Mice are fixed by cardiac perfusion using a saline rinse followed by 4% paraformaldehyde (PFA). After the tissues are dissected and rinsed in phosphate buffered saline (PBS), the tissues are post-fixed in 4% PFA for 30 minutes. Tissues are then rinsed in PBS and soaked overnight in first 15% and then 30% sucrose. The tissues are trimmed and frozen in cryo-protectant (OCT). The embedded tissue blocks are sectioned between 10 and 14 microns and then the sections are stained in lacZ staining solution for 18 hours. Slides are then post-fixed in 10% neutral buffered formalin (NBF), rinsed in PBS, counterstained using Nuclear Fast Red, dehydrated in an ethanol series, cleared using Xylene, and coverslipped with a Xylene based mounting media. Slides are scanned using an Imperio ScanScope XT Slide Scanner and viewed with Aperio Image Scope software, version 10.2.2.2319. All lacZ staining is photodocumented and noted as part of the annotation information.

Lexicon Genentech Phenotyping Data

Data and germplasm from 472 knockout mice were recently donated by Lexicon & Genentech Pharmaceuticals and are publicly available through the KOMP-Phenotyping website. Phenotyping data were collected from pedigrees containing wildtype, heterozygous and homozygous mutants. The phenotyping data are comprehensive and include: serum chemistry; blood pressure and heart rate; organ & body weights and body length; hematology and blood FACS analysis; imaging with CAT, DEXA & microCT; a behavioral test battery; skin proliferation assay; eye angiogram & funduscope; and gross & microscopic pathology. For a recent publication describing these animals and associated data see: Tang et al., Nature Biotechnology 28(7): 749-755, 2010. View the Lexicon Genentech gene list.