Shc1fl/fl floxed mice may be useful for studying SHC1 requirements during development, T cell maturation, and tumorigenesis.
Kodi S. Ravichandran, University of Virginia
Shc1fl/fl floxed mutant mice possess loxP sites flanking exons 1-3 of the src homology 2 domain-containing transforming protein C1 (Shc1) gene. SHC1 acts downstream of many different receptors including growth factor receptors, integrins, TCR, BCR, and the preTCR. Mice that are homozygous for the floxed Shc1 allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exons 1-3, including the transcriptional start site and the amino terminals of the three known Shc1 isoforms, deleted in cre-expressing tissues. Breeding these mice to mice with widespread cre expression results in embryonically lethality by E10.5.
For example, when crossed to a strain expressing Cre recombinase in thymocytes (see Stock No. 012837 ), resulting mice exhibit a block at the beta selection checkpoint and have greatly reduced numbers of double positive and single positive thymocytes, and an increase in percentage of CD4-CD8- DN thymocytes.
A targeting vector was designed to insert a single loxP site upstream of exon 1, and a loxP-flanked neomycin resistance (neo) cassette downstream of exon 2 of the src homology 2 domain-containing transforming protein C1 (Shc1) gene. The construct was electroporated into 129Sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a pIC-Cre expression plasmid to delete the neo cassette. Resulting ES cells contained multiple gene rearrangments; intact floxed-exons, intact floxed-neo cassette, or excision of floxed exons and the neo cassette. Correctly targeted ES cells, containing only the floxed-exons, were injected into C57BL/6 blastocysts and resulting chimeric males were bred with C57BL/6J females. Resulting offspring were bred to C57BL/6J mice for at least 10 to establish a colony of Shc1fl/fl mice. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
|Allele Name||targeted mutation 1, Kodi S Ravichandran|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Shc1, src homology 2 domain-containing transforming protein C1|
|Strain of Origin||129|
|Molecular Note||Exons 1 and 2, encoding the transcriptional start site and the amino terminals of the three known isoforms, were flanked by a single upstream loxP site and a floxed neo cassette inserted into intron 2. Transient transfection of ES cells resulted in the excision of the floxed neo cassette, leaving the first two exons surrounded by single loxP sites.|
When maintaining a live colony, homozygous mice may be bred together.
When using the B6.129-Shc1tm1Ravi/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #025879 in your Materials and Methods section.