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Sci. Rep. 16, 4694. Imaging of conditional gene silencing in vivo using a bioluminescence-based method with thermo-inducible microRNAs 2018

Pinel, K., Genevois, C., Debeissat, C., Couillaud, F.

Notes: A novel therapeutic method utilizing synthetic microRNAs combined with a heat shock-inducible promoter to decrease target gene expression is described. This method is ideal for diseases where target gene overexpression leads to disease state. To monitor gene expression, the firefly luciferase gene within a tumor is targeted and expression is monitored using the Dual-Luciferase® Reporter Assay System. The ViviRen™ Live Cell Substrate is used to monitor expression in live mice. (5088)

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J. Biol. Chem. 292, 10651–63. A split-luciferase complementation, real-time reporting assay enables monitoring of the disease-associated transmembrane protein TREM2 in live cells. 2017

Varnum, M.M., Clayton, K.A., Yoshii-Kitahara, A., Yonemoto, G., Koro, L. Ikezu, S. and Ikezu, T.

Notes: Triggered receptor expressed on myeloid cells 2 (TREM2) is associated with microglial inflammation and mutations are linked to various neurodegenerative diseases. A split Renilla luciferase system was used to investigate the TREM2 and TYPRO protein-tyrosine kinase-binding protein (TYROBP) interaction. HEK293 cells were treated with anti-human TREM2 antibody, leading to stimulation of TREM2 dimerization and TYROBP coupling. Coupling led to a significant increase in luminescence. Together, a system for monitoring protein:protein interactions using a split Renilla luciferase is demonstrated. (5114)

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J. Exp. Bot. 38, 4171–83. Heterodimerization of Arabidopsis calcium/proton exchangers contributes to regulation of guard cell dynamics and plant defense responses. 2017

Hocking, B., Conn, S.J., Manohar, M., Xu, B., Athman, A., Stancombe, M.A., Webb, A.R., Hirschi, K.D. and Gilliham, M.

Notes: The multimerization of cation exchangers (CAX1 and CAX3) was investigated in Arabidopsis thaliana. A split luciferase assay using the ViviRen™ Live Cell Substrate demonstrated the formation of both homo- and heterodimers of CAX1 and CAX3. CAX1 knockout strains and strains exposed to pathogens showed an increase in CAX3 expression. CAX promoter luciferase fusions demonstrated an increase in promoter activity under these conditions. (5113)

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Proc. Natl. Acad. Sci. USA 104, 10264-10269. Imaging protein interactions with bioluminescence resonance energy transfer (BRET) in plant and mammalian cells and tissues. 2007

Xu, X., Soutto, M., Xie, Q., Servick, S., Subramanian, C., von Arnim, A.G., Johnson, C.H.

Notes: These authors used the coelenterazine analog ViviRen® as a substrate for Renilla luciferase in Bioluminescence Resonance Energy Transfer (BRET) experiments. Using this technique, they were able to demonstrate protein interactions in single mammalian cells. (3631)

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