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Functional imaging of Rel expression in inflammatory processes using bioluminescence imaging system in transgenic mice.

Yang X, Jing H, Zhao K, Sun R, Liu Z, Ying Y, Ci L, Kuang Y, Huang F, Wang Z, Fei J - PLoS ONE (2013)

Bottom Line: Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs.Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model.These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Technology, Tongji University, Shanghai, China.

ABSTRACT
c-Rel plays important roles in many inflammatory diseases. Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs. In this paper, a transgenic mouse line, B6-Tg(c-Rel-luc)(Mlit), which incorporated the transgene firefly luciferase driven by a 14.5-kb fragment containing mouse c-Rel gene Rel promoter, was generated to monitor Rel expression in vivo. Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model. The luciferase expression in transgenic mice was comparable to the endogenous Rel expression and could be suppressed by administration of anti-inflammatory drug dexamethasone or aspirin. These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

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Schematic diagram of pc-Rel-luc reporter construct and identification of the transgenic founders.(A) The c-Rel-luc transgene harboring a 14.6 kb mouse Rel gene promoter and firefly luciferase cDNA was diagramed. Primer P1 and P2 was indicated. P1: forward-luc primer, P2: reverse-luc primer. (B) A 604-bp fragment was amplified by PCR with primer P1 and P2 in transgenic founder mice. PCR products were separated on 1% agarose gel. Lane 1–9: the products from transgenic founders; lane 10: from a wild-type C57BL/6 control; lane 11: from ddH2O; lane 12: DNA marker DL 2,000 ladder.
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pone-0057632-g001: Schematic diagram of pc-Rel-luc reporter construct and identification of the transgenic founders.(A) The c-Rel-luc transgene harboring a 14.6 kb mouse Rel gene promoter and firefly luciferase cDNA was diagramed. Primer P1 and P2 was indicated. P1: forward-luc primer, P2: reverse-luc primer. (B) A 604-bp fragment was amplified by PCR with primer P1 and P2 in transgenic founder mice. PCR products were separated on 1% agarose gel. Lane 1–9: the products from transgenic founders; lane 10: from a wild-type C57BL/6 control; lane 11: from ddH2O; lane 12: DNA marker DL 2,000 ladder.

Mentions: The schematic diagram of c-Rel-luc transgene is showed in Fig. 1A. The 14567 bp DNA fragment upstream from the translational initial codon ATG of mouse Rel gene was used as the promoter to drive the expression of firefly luciferase. The PCR primers designed for identification of the transgene were located in the region of Rel sequence and the luciferase gene (Fig. 1A). Nine founders were obtained (Fig. 1B) and the transgenic mice, here named as B6-Tg(c-Rel-luc)Mlit, were backcrossed to wild type C57BL/6J mice at least for five generations for further studies.


Functional imaging of Rel expression in inflammatory processes using bioluminescence imaging system in transgenic mice.

Yang X, Jing H, Zhao K, Sun R, Liu Z, Ying Y, Ci L, Kuang Y, Huang F, Wang Z, Fei J - PLoS ONE (2013)

Schematic diagram of pc-Rel-luc reporter construct and identification of the transgenic founders.(A) The c-Rel-luc transgene harboring a 14.6 kb mouse Rel gene promoter and firefly luciferase cDNA was diagramed. Primer P1 and P2 was indicated. P1: forward-luc primer, P2: reverse-luc primer. (B) A 604-bp fragment was amplified by PCR with primer P1 and P2 in transgenic founder mice. PCR products were separated on 1% agarose gel. Lane 1–9: the products from transgenic founders; lane 10: from a wild-type C57BL/6 control; lane 11: from ddH2O; lane 12: DNA marker DL 2,000 ladder.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585201&req=5

pone-0057632-g001: Schematic diagram of pc-Rel-luc reporter construct and identification of the transgenic founders.(A) The c-Rel-luc transgene harboring a 14.6 kb mouse Rel gene promoter and firefly luciferase cDNA was diagramed. Primer P1 and P2 was indicated. P1: forward-luc primer, P2: reverse-luc primer. (B) A 604-bp fragment was amplified by PCR with primer P1 and P2 in transgenic founder mice. PCR products were separated on 1% agarose gel. Lane 1–9: the products from transgenic founders; lane 10: from a wild-type C57BL/6 control; lane 11: from ddH2O; lane 12: DNA marker DL 2,000 ladder.
Mentions: The schematic diagram of c-Rel-luc transgene is showed in Fig. 1A. The 14567 bp DNA fragment upstream from the translational initial codon ATG of mouse Rel gene was used as the promoter to drive the expression of firefly luciferase. The PCR primers designed for identification of the transgene were located in the region of Rel sequence and the luciferase gene (Fig. 1A). Nine founders were obtained (Fig. 1B) and the transgenic mice, here named as B6-Tg(c-Rel-luc)Mlit, were backcrossed to wild type C57BL/6J mice at least for five generations for further studies.

Bottom Line: Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs.Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model.These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Technology, Tongji University, Shanghai, China.

ABSTRACT
c-Rel plays important roles in many inflammatory diseases. Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs. In this paper, a transgenic mouse line, B6-Tg(c-Rel-luc)(Mlit), which incorporated the transgene firefly luciferase driven by a 14.5-kb fragment containing mouse c-Rel gene Rel promoter, was generated to monitor Rel expression in vivo. Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model. The luciferase expression in transgenic mice was comparable to the endogenous Rel expression and could be suppressed by administration of anti-inflammatory drug dexamethasone or aspirin. These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

Show MeSH
Related in: MedlinePlus