Green fluorescent protein as a marker for gene expression

Chalfie, Martin

doi:10.1016/0168-9525(94)90088-4

Cited by 1,557 publications

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“…The other reporter gene construct encoding green fluorescent protein has the added advantage that as well as being detectable in single cells it does not require fixative and hence can be used in living cells. 39 However, some tissues, including the heart have high levels of background autofluorescence.…”

Section: Discussionmentioning

confidence: 99%

β-Galactosidase staining following intracoronary infusion of cationic liposomes in the in vivo rabbit heart is produced by microinfarction rather than effective gene transfer: a cautionary tale

et al. 1998

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The myocardium is a potential target for the expression of conversion at day 3 with associated myocardial damage. exogenous genes to treat inherited and acquired diseases.We hypothesised that the damage was associated with Although adenovirus-mediated gene transfer has resulted macro-aggregates of cationic liposomes-DNA occluding in high-level gene transfer in vivo via direct intramyocardial the microcirculation. When such aggregates were excluded injection and via a percutaneous intra-arterial route, the no X-gal conversion was seen in vivo. In order to show time-course of gene expression is limited by host immune that X-gal conversion occurs in areas of infarction in the responses. It was the aim of this study to test whether catmyocardium we caused closed chest infarction by ionic liposome-mediated gene transfer, which does not sufdeploying a platinum micro-embolisation coil in the circumfer from the aforementioned problems, was feasible in the flex coronary artery. At day 3 X-gal conversion was adult rabbit myocardium via a percutaneous transluminal observed in the territory supplied by the occluded artery. approach. Doses of plasmid DNA encoding lacZ from 200-Thus, microinfarction causes the false positive appearance 800 g complexed to cationic liposomes resulted in X-gal of gene transfer when using a lacZ reporter gene.

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Section: Discussionmentioning

confidence: 99%

β-Galactosidase staining following intracoronary infusion of cationic liposomes in the in vivo rabbit heart is produced by microinfarction rather than effective gene transfer: a cautionary tale

et al. 1998

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“…Genes transferred by rAAV carrying the cytomegalovirus (CMV) immediate-early promoter have been found to be expressed preferentially in neurons. 10 In this study, we explored the possibility of gene expression in oligodendrocytes mediated by rAAV, using the green fluorescent protein (GFP) 11,12 as a reporter gene under the control of the myelin basic protein (MBP) transcriptional control region. 13 …”

Section: Introductionmentioning

confidence: 99%

Gene transfer and expression in oligodendrocytes under the control of myelin basic protein transcriptional control region mediated by adeno-associated virus

et al. 1998

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In this study, a rAAV vector carrying a reporter gene,Infusion of approximately 6 × 10 9 particles (2 × 10 5 infec-'humanized' green fluorescent protein (GFP), linked to the tious units) of rAAV-MBP-GFP into mouse brains resulted transcriptional control region from the myelin basic protein in the GFP expression specifically in white matter. The (MBP) gene (a myelin-forming cell-specific gene) was con-GFP protein was detected 15 days later by immunostainstructed. Transduction of oligodendrocytes was carried out ing, specifically in the oligodendrocytes. No astrocytes both in vitro and in vivo. The GFP expression was detected were transduced. Our studies suggested that cell types for at least 3 weeks in both transduced oligodendrocyte cell other than neurons in the central nervous system can also line (MOCH-1 cells) and primary cultures of rat oligodendbe transduced by rAAV using a cell-type-specific transcriprocytes. Preferential GFP expression in oligodendrocytes tional control region or promoter. The MBP transcriptional was observed in the primary cultures. In contrast, transduccontrol region might be suitable for gene therapy and other tion with rAAV carrying the CMV promoter produced neurobiology studies requiring direct targeting to the myelistronger GFP fluorescence in various cell types, with the nating cells. majority of GFP-expressing cells being the astrocytes.

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“…The GFP emits bright green light after exposure to ultraviolet or blue light without extrinsic labeling or substrates. With the wild-type (wt) GFP gene being cloned, it has been used as a reporter gene in a real-time and noninvasive fashion (Chalfie et al, 1994). Recently, variants of GFP have also been designed in that they are adapted better to mammalian expression and signal detection.…”

Section: Introductionmentioning

confidence: 99%

Rapid diagnosis and quantification of herpes simplex virus with a green fluorescent protein reporter system

Kung¹,

Wang

Lin³

et al. 2000

Journal of Virological Methods

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A genetically modified cell line (Vero-ICP10-EGFP) was constructed for detection of herpes simplex virus (HSV) by a simple, rapid and direct method. The cell line was developed by stable transfection of Vero cell with a plasmid encoding the green fluorescent protein (GFP) driven by the promoter of the HSV-2 ICP10 gene. As early as 6 h after infection with HSV, fluorescence-emitting cells can be observed under a fluorescence microscope. A single infected cell emitting fluorescence can be observed with soft agar overlay by inverted fluorescence microscopy. No induction of detectable fluorescence was seen following infections with human cytomegalovirus (HCMV), varicella zoster virus (VZV), coxsackievirus A16 and enterovirus 71. Analysis by flow cytometry also demonstrated that intensity of the triggered fluorescence is proportional to the titer of HSV inoculated. Taken together, this novel GFP reporter system could become a useful means for rapid detection and quantification of HSV in clinical specimens.

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Green fluorescent protein as a marker for gene expression

Cited by 1,557 publications

References 0 publications

β-Galactosidase staining following intracoronary infusion of cationic liposomes in the in vivo rabbit heart is produced by microinfarction rather than effective gene transfer: a cautionary tale

β-Galactosidase staining following intracoronary infusion of cationic liposomes in the in vivo rabbit heart is produced by microinfarction rather than effective gene transfer: a cautionary tale

Gene transfer and expression in oligodendrocytes under the control of myelin basic protein transcriptional control region mediated by adeno-associated virus

Rapid diagnosis and quantification of herpes simplex virus with a green fluorescent protein reporter system

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