Tetrameric far‐red fluorescent protein as a scaffold to assemble an octavalent peptide nanoprobe for enhanced tumor targeting and intracellular uptake in vivo

2011 ◽  
Vol 25 (6) ◽  
pp. 1865-1873 ◽  
Author(s):  
Haiming Luo ◽  
Jie Yang ◽  
Honglin Jin ◽  
Chuan Huang ◽  
Jianwei Fu ◽  
...  
Keyword(s):  
Tumor Targeting ◽  
Fluorescent Protein ◽  
Intracellular Uptake ◽  
Red Fluorescent Protein

Fluorescent proteins for in vivo imaging, where's the biliverdin?

2020 ◽  
Vol 48 (6) ◽  
pp. 2657-2667
Author(s):  
Felipe Montecinos-Franjola ◽  
John Y. Lin ◽  
Erik A. Rodriguez
Keyword(s):  
Hydrogen Peroxide ◽  
In Vivo Imaging ◽  
Near Infrared ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Fluorescent Imaging ◽  
Infrared Light ◽  
Red Fluorescent Protein ◽  
Color Palette

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light >600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10−18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo.

scholarly journals Small ultra-red fluorescent protein nanoparticles as exogenous probes for noninvasive tumor imaging in vivo

2020 ◽  
Vol 153 ◽  
pp. 100-106 ◽  
Author(s):  
Feifei An ◽  
Nandi Chen ◽  
William J. Conlon ◽  
Justin S. Hachey ◽  
Jingqi Xin ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Tumor Imaging ◽  
Red Fluorescent Protein ◽  
Protein Nanoparticles

Testicular injection of busulfan for recipient preparation in transplantation of spermatogonial stem cells in mice

2016 ◽  
Vol 28 (12) ◽  
pp. 1916 ◽  
Author(s):  
Yusheng Qin ◽  
Ling Liu ◽  
Yanan He ◽  
Wenzhi Ma ◽  
Huabin Zhu ◽  
...  
Keyword(s):  
Germ Cells ◽  
Fluorescent Protein ◽  
Positive Control ◽  
Negative Control ◽  
Seminiferous Tubules ◽  
Red Fluorescent Protein ◽  
Efferent Duct ◽  
Testicular Injection ◽  
Transplantation Recipient

Intraperitoneal busulfan injections are used to prepare recipients for spermatogonial stem cell (SSC) transplantation but they are associated with haematopoietic toxicity. Testicular injections of busulfan have been proposed to overcome this limitation. To date, testicular injections have not been studied in the mouse model. Therefore, in the present study we used ICR mice as recipients for SSC transplantation and prepared these mice by testicular injection of busulfan on both sides (2, 3, 4 or 6 mg kg–1 per side). Following this, donor germ cells expressing red fluorescent protein (RFP) from transgenic C57BL/6J male mice were transplanted into recipients via the efferent duct on Days 16–17 after busulfan treatment. Positive control mice were prepared by intraperitoneal injection of 40 mg kg–1 busulfan and negative control mice were treated with bilateral testicular injection of 50% dimethyl sulfoxide. On Day 49 after transplantation, recipient mice that were RFP-positive by in vivo imaging were mated with ICR female mice. Donor-derived germ cell colonies with red fluorescence were observed on Day 60 after transplantation, and donor-derived offspring were obtained. The results demonstrated that endogenous germ cells were successfully eliminated in the seminiferous tubules via testicular busulfan administration, and that exogenous SSCs successfully undergo spermatogenesis in the testes of recipient mice prepared by testicular injections of busulfan. In addition to its effects on recipient preparation, this method was safe in rodents and could possibly be adapted for use in other species.

scholarly journals Rational design of a monomeric and photostable far-red fluorescent protein for fluorescence imaging in vivo

2015 ◽  
Vol 25 (2) ◽  
pp. 308-315 ◽  
Author(s):  
Dan Yu ◽  
Zhiqiang Dong ◽  
William Clay Gustafson ◽  
Rubén Ruiz-González ◽  
Luca Signor ◽  
...  
Keyword(s):  
Fluorescence Imaging ◽  
Rational Design ◽  
Fluorescent Protein ◽  
Red Fluorescent Protein

Platelets release mitochondrial antigens in systemic lupus erythematosus

2021 ◽  
Vol 13 (581) ◽  
pp. eaav5928 ◽  
Author(s):  
Imene Melki ◽  
Isabelle Allaeys ◽  
Nicolas Tessandier ◽  
Tania Lévesque ◽  
Nathalie Cloutier ◽  
...  
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Fluorescent Protein ◽  
Red Fluorescent Protein ◽  
Systemic Lupus ◽  
Fibrinogen Receptor ◽  
Mitochondrial Antigens ◽  
Stimulation Of

The accumulation of DNA and nuclear components in blood and their recognition by autoantibodies play a central role in the pathophysiology of systemic lupus erythematosus (SLE). Despite the efforts, the sources of circulating autoantigens in SLE are still unclear. Here, we show that in SLE, platelets release mitochondrial DNA, the majority of which is associated with the extracellular mitochondrial organelle. Mitochondrial release in patients with SLE correlates with platelet degranulation. This process requires the stimulation of platelet FcγRIIA, a receptor for immune complexes. Because mice lack FcγRIIA and murine platelets are completely devoid of receptor capable of binding IgG-containing immune complexes, we used transgenic mice expressing FcγRIIA for our in vivo investigations. FcγRIIA expression in lupus-prone mice led to the recruitment of platelets in kidneys and to the release of mitochondria in vivo. Using a reporter mouse with red fluorescent protein targeted to the mitochondrion, we confirmed platelets as a source of extracellular mitochondria driven by FcγRIIA and its cosignaling by the fibrinogen receptor α2bβ3 in vivo. These findings suggest that platelets might be a key source of mitochondrial antigens in SLE and might be a therapeutic target for treating SLE.

scholarly journals Use of the mCherry Fluorescent Protein To Study Intestinal Colonization by Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 in Mice

2015 ◽  
Vol 81 (17) ◽  
pp. 5993-6002 ◽  
Author(s):  
Winschau F. van Zyl ◽  
Shelly M. Deane ◽  
Leon M. T. Dicks
Keyword(s):  
Lactobacillus Plantarum ◽  
Fluorescent Protein ◽  
Intestinal Colonization ◽  
Red Fluorescent Protein ◽  
Gene Encoding ◽  
Content Type ◽  
Enterococcus Mundtii ◽  
Probiotic Strains ◽  
Reporter Systems

ABSTRACTLactic acid bacteria (LAB) are natural inhabitants of the gastrointestinal tract (GIT) of humans and animals, and some LAB species receive considerable attention due to their health benefits. Although many papers have been published on probiotic LAB, only a few reports have been published on the migration and colonization of the cells in the GIT. This is due mostly to the lack of efficient reporter systems. In this study, we report on the application of the fluorescent mCherry protein in thein vivotagging of the probiotic strainsEnterococcus mundtiiST4SA andLactobacillus plantarum423. ThemCherrygene, encoding a red fluorescent protein (RFP), was integrated into a nonfunctional region on the genome ofL. plantarum423 by homologous recombination. In the case ofE. mundtiiST4SA, themCherrygene was cloned into the pGKV223D LAB/Escherichia coliexpression vector. Expression of themCherrygene did not alter the growth rate of the two strains and had no effect on bacteriocin production. Both strains colonized the cecum and colon of mice.

scholarly journals Illuminating the Formation of Lumens

2007 ◽  
Vol 15 (3) ◽  
pp. 3-5
Author(s):  
Stephen W. Carmichael
Keyword(s):  
Animal Model ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Solid Core ◽  
Red Fluorescent Protein ◽  
Neuronal Process ◽  
Two Photon ◽  
Green Fluorescent

How do lumens form? Two mechanisms that come readily to mind are a wrapping model, similar to the wrapping of the myelin sheath around a neuronal process, and a solid core of cells followed by apoptosis of the central cells. Another obvious mechanism that was suggested over 100 years ago is the fusion of intracellular vacuoles. Whereas several recent studies have supported this latter mechanism, it has not yet been proven. Now, the appropriate animal model (zebrafish), the modern techniques (transgenic chimeras), dyes (green fluorescent protein and monomeric red fluorescent protein) that can be linked to proteins to label vacuoles, and two-photon imaging in real time finally have provided the strongest support yet. In an article by Makoto Kamei, Brian Saunders, Kayla Bayless, Louis Dye, George Davis, and Brant Weinstein the assembly of endothelial tubes from intracellular vacuoles was observed in vitro and in vivo.

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