Mouse embryonic fibroblasts (MEFs) from mice homozygous for this targeted mutation exhibit impaired migration, cell spreading, transferrin recycling and reduced cholesterol levels. These mice may be useful in studies of endocytic transport, intracellular trafficking and recycling of membrane proteins, and cholesterol homeostasis.
Mia Horowitz, Tel Aviv University
The protein encoded by the Ehd1 (EH-domain containing 1) gene contains a protein-binding EH (EPS15 homology) domain and has a role in early endocytic membrane fusion and membrane trafficking of recycling endosomes. Mice that are homozygous for this targeted mutation are viable and fertile. No gene product (protein) is detected by Western blot analysis of lung, heart, testis and skeletal muscle from homozygous mice. However, a truncated 64kDa protein is detected by Western blot analysis of brain tissue from homozygous mice. No gene product (mRNA) is detected in primary mouse embryonic fibroblasts (MEFs) from homozygotes by RT-PCR. Ehd1 deficient MEFs plated on fibronectin have impaired migration and cell spreading. MEFs from homozygotes also exhibit impaired transferrin recycling, lower overall levels of beta-1 integrins on the plasma membrane, abnormal focal adhesion formation, and lower cellular cholesterol and reduced lipid droplet size.
A targeting vector designed by Dr. Mia Horowitz (Tel Aviv University) containing a NEO cassette was inserted into exon 3, which resulted in disruption of part of exon 3, and all of exons 4 and 5. The construct was electroporated into 129S6/SvEvTac derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were aggregated with morulae. The resulting chimeric males were bred to 129/SvEv or Swiss Webster females. The mice were then bred to ICR. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
|Allele Name||targeted mutation 1, Mia Horowitz|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Ehd1, EH-domain containing 1|
|Strain of Origin||129S6/SvEvTac|
|Molecular Note||A neomycin resistance cassette was inserted into exon 3 such that it fused to the first 164 bp of that exon and replaced exons 4 and 5.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the STOCK Ehd1tm1Mhor/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #023073 in your Materials and Methods section.