A one-plasmid conditional color-switching transgenic system for multimodal bioimaging.
We have developed a new construct to generate transgenic mice with one plasmid that offers: (1) Cre/loxP-mediated spatial and temporally-controlled tissue-specific transgene expression; (2) A color-switching mechanism that uses spectrum-complementary genetically-encoded red (mRFP) and green (eGFP) fluorescent markers to label the transgene-expressing cells; (3) A bioluminescent marker that turns-on in the transgene-expressing cells; (4) eGFP as a cell surface marker in the transgene-expressing cells that facilitates the isolation and targeting of these cells. This vector was tested in vitro by co-transfection of the transgenic plasmid and a plasmid containing Cre recombinase into cultured cells and by establishing a transgenic mouse line. We show that this method allows versatile transgene expression targeting and color-switching to facilitate fluorescent and bioluminescent imaging both in cultured cells and in vivo. Our strategy provides time-saving features in tissue-specific transgene expression, bioimaging and primary cell isolation and can be used for generation of gene-specific transgenic mice.
Zheng
Lei
L
Department of Cell Biology and Neuroscience, University of California-Riverside, Riverside, CA 92521, USA.
Njauw
Ching-Ni
CN
Martins-Green
Manuela
M
D000818:Animals; D000831:Animals, Newborn / Q000502:physiology*; D004965:Estrogen Antagonists / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D049449:Luminescence; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D041681:NIH 3T3 Cells; D010641:Phenotype; D010957:Plasmids; D011506:Proteins / Q000502:physiology; D022661:RNA, Untranslated; D011960:Receptors, Estrogen / Q000502:physiology; D013629:Tamoxifen / Q000494:pharmacology; D014018:Tissue Distribution; D000097573:Red Fluorescent Protein
D004965:Estrogen Antagonists; D049452:Green Fluorescent Proteins; D019426:Integrases; D008156:Luciferases; D008164:Luminescent Proteins; D011506:Proteins; D022661:RNA, Untranslated; D011960:Receptors, Estrogen; D013629:Tamoxifen; D000097573:Red Fluorescent Protein; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; C485184:enhanced green fluorescent protein; C045073:Cre recombinase