PI3Kγ deficient mice express GFP in hematopoietic cells, muscle, and the pancreas and may be useful when studying the role of p110γ in inflammation and inflammatory diseases.
Dianqing Wu, Yale University School of Medicine
A green fluorescent protein (GFP) and a neomycin resistance (neo) cassette replaces part of exon 1 and exons 2-3 of the phosphoinositide-3-kinase, catalytic, gamma polypeptide (Pik3cg) gene, abolishing gene expression. PI3Kγ, p110γ, is expressed in hematopoietic cells, muscle, and the pancreas and facilitates the migration of leukocytes, including neutrophils and eosinophils, to the site of inflammation. Homozygous are viable and fertile. GFP is detected in the splenocytes, bone marrow cells, and neutrophils. GFP was detected with flow cytometry in over 90% of Mac1+ cells in the peritoneum, and 82% of CD45R+ and 70% of CD3+ cells from the spleens of PI3Kγ deficient mice. These mice display impaired neutrophil chemotaxis and respiratory burst in response to formyl peptide N-formyl-Met-Leu-Phe (fMLP) and C5a induction, as well as reduced thymocyte survival and activation of mature T lymphocytes. Neutrophils have decreased PIP3 production and AKT activation, and peritoneal macrophages display reduced migration and accumulation in a septic peritonitis model. These PI3Kγ deficient mice exhibit decreased bone reabsorption and an increase in apoptosis during osteoclast differentiation. An increase in IL-17A production in also seen in splenocytes.
A targeting vector was designed to replace part of exon 1 and exons 2-3 of the phosphoinositide-3-kinase, catalytic, gamma polypeptide (Pik3cg) gene with a green fluorescent protein (GFP) and a neomycin resistance (neo). The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl+-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to C57BL/6NHsd females. These mice were bred to C57BL/6NHsd for at least 12 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ (Stock No. 005304) for at least one generation to establish the colony.
|Expressed Gene||GFP, Green Fluorescent Protein, jellyfish|
|Site of Expression|
|Allele Name||targeted mutation 1, Dianqing Wu|
|Allele Type||Targeted (Null/Knockout, Reporter)|
|Allele Synonym(s)||PI3Kgamma-; p110gamma-|
|Gene Symbol and Name||Pik3cg, phosphoinositide-3-kinase, catalytic, gamma polypeptide|
|Gene Synonym(s)||5830428L06Rik; 5830428L06Rik; PI(3)Kgamma; PI3CG; PI3K; PI3Kgamma; PI3Kgamma; PIK3; Pi3k; RIKEN cDNA 5830428L06 gene; p110gamma; p120-PI3K|
|Expressed Gene||GFP, Green Fluorescent Protein, jellyfish|
|Strain of Origin||Not Specified|
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