A hCXCR1 transgenic mouse model containing a conditional color-switching system for imaging of hCXCL8/IL-8 functions in vivo.
To address the functions of human CXCL8 (hCXCL8)/IL-8 through hCXCR1 in vivo, we have developed a humanized, transgenic mouse for hCXCR1. This mouse line is versatile and allows for a variety of functional analyses using bioimaging, including Cre/loxP-mediated, tissue-specific hCXCR1 expression in a spatiotemporal manner; a color-switching mechanism, which uses spectrum-complementary, genetically encoded green and red fluorescence markers to label the hCXCR1-expressing cells [enhanced GFP (eGFP)] against the background [monomeric red fluorescent protein (mRFP)]; a bioluminescent marker, which is present in the hCXCR1-expressing cells; and an exogenous cell surface marker (eGFP moiety) in the hCXCR1-expressing cells, which facilitates identification, isolation, and targeting of these cells. The established, transgenic founder line RCLG3A (TG(+)) expresses only mRFP and does so ubiquitously. When the RCLG3A mice are crossed with the tamoxifen-inducible, whole-tissue Cre mice (ROSA26-Cre/Esr(+/-)), administration of tamoxifen induces whole-body hCXCR1 expression and color-switching. When RCLG3A mice are crossed with thymocyte-specific Cre mice (Lck-Cre(+/+)), the hCXCR1 expression and color-switching are restricted in a lineage-specific manner. This mouse line can be used to understand the functions of hCXCL-8 in vivo. In addition, our approach and vectors can be used to establish other tissue-specific, transgenic mice in conjunction with multifunctional cell markers, which facilitate cell imaging, tracing, and manipulation in vivo.
Zheng
Lei
L
Department of Cell Biology and Neurosciences, University of California Riverside, Riverside, CA 92521, USA.
Njauw
Ching-Ni
CN
Martins-Green
Manuela
M
D000818:Animals; D002633:Chemotaxis; D004965:Estrogen Antagonists / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D019426:Integrases; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D041681:NIH 3T3 Cells; D010641:Phenotype; D023062:Receptors, Interleukin-8A / Q000235:genetics / Q000502:physiology*; D012867:Skin / Q000166:cytology / Q000378:metabolism; D013629:Tamoxifen / Q000494:pharmacology*; D000097573:Red Fluorescent Protein
D004965:Estrogen Antagonists; D049452:Green Fluorescent Proteins; D019426:Integrases; D016209:Interleukin-8; D008156:Luciferases; D008164:Luminescent Proteins; D023062:Receptors, Interleukin-8A; D013629:Tamoxifen; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein; C045073:Cre recombinase