Vcam1tm2Dmil
Targeted Allele Detail
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Symbol: |
Vcam1tm2Dmil |
Name: |
vascular cell adhesion molecule 1; targeted mutation 2, David Milstone |
MGI ID: |
MGI:5700541 |
Synonyms: |
VcamkB |
Gene: |
Vcam1 Location: Chr3:115903669-115923337 bp, - strand Genetic Position: Chr3, 50.17 cM
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Alliance: |
Vcam1tm2Dmil page
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Germline Transmission: |
Earliest citation of germline transmission:
J:227109
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Parent Cell Line: |
J1 (ES Cell)
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Strain of Origin: |
129S4/SvJae
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Allele Type: |
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Targeted (Not Applicable) |
Mutations: |
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Insertion, Nucleotide substitutions
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Mutation details: Sequential tag and exchange gene targeting was used to introduce a 6 nucleotide mutation in the 5NF-B binding site of the transcriptional promoter, and a 3 nucleotide mutation in the 5UTR of Vcam1 in exon 1. The first, "tag", targeting vector replaces exon 1 (encoding the signal peptide) and part of exon 2 (encoding immunoglobulin-like domain 1) with a pgk-neomycin/TK cassettes. The Tag construct is transfected into embryonic stems cells and a single correctly targeted clone is transfected with the second construct.
The second "Exchange" targeting construct replaces the neo cassette and introduces a 6 nucleotide substitution 5' of the transcription start site resulting in the disruption of the 5'NFKB1 (NF-kappaB) binding site domain and a 3 nucleotide substitution downstream of the translation start site in the 5' UTR of exon 1.
The 3 nt substitution is used to distinguish the mutant and wild-type mRNA and hnRNA. Western blot ananlysis demonstrates the presence of mutant protein in heart endothelial cells.
(J:227109)
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View phenotypes and curated references for all genotypes (concatenated display).
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Original: |
J:227109 Milstone DS, et al., Differential role of an NF-kappaB transcriptional response element in endothelial versus intimal cell VCAM-1 expression. Circ Res. 2015 Jul 3;117(2):166-77 |
All: |
1 reference(s) |
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