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GFP adenoviral particles

GFP adenoviral particles

Cat.No. :  AD00281Z

Titer: ≥1x10^10 IFU/mL / ≥1x10^11 IFU/mL / ≥1x10^11 VP/mL / ≥1x10^12 VP/mL Size: 100 ul/500 ul/1 mL

Storage:  -80℃ Shipping:  Frozen on dry ice

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Adenovirus Particle Information

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Cat. No. AD00281Z
Description This recombinant human adenovirus type 5 expresses enhanced green fluorescent protein under the control of a CMV promoter. Green fluorescent protein (GFP) is a protein originally isolated from the jellyfish that fluoresces green when exposed to light in the blue to ultraviolet range.
Target Gene gfp
Product Type Adenoviral particle
Insert GFP
Titer Varies lot by lot, for example, ≥1x10^10 IFU/mL, ≥1x10^11 IFU/mL, ≥1x10^11 VP/mL etc.
Size Varies lot by lot, for example, 250 ul, 500 ul, 1 mL etc.
Storage Store at -80℃. Avoid multiple freeze/thaw cycles.
Shipping Frozen on dry ice
Creative Biogene ensures high-quality adenovirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between adenovirus particle lots.
Endotoxin Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in adenovirus production, especially for applications in animal studies and gene therapy. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced adenovirus particles to ensure regulatory compliance.
Sterility Creative Biogene ensures that adenovirus products are free of any bacterial, fungal and other microbial contamination.
Ad5 E1 Detection All Creative Biogene adenoviruses are PCR tested to ensure that there are no detectable E1 sequences in the particles, which could be from revertants or external E1 contamination.
RCA Assays Adenovirus products originating at Creative Biogene are guaranteed to have undetectable replication-competent adenovirus (RCA). This quality control measure is important because there is always the possibility of wild-type contamination due to revertants or environmental sources.
PFU Titering All purified adenovirus preparations are tested for infectious titer. Creative Biogene's PFU test takes a few days longer but counts true plaques in HEK cells rather than estimating PFU titers via IHC staining or TCI50 of infected cells.
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Human GFP adenoviral particles are recombinant viral vectors genetically engineered to deliver and express the green fluorescent protein (GFP) gene in human or mammalian cells. These particles are derived from a human adenovirus, specifically adenovirus type 5 (Ad5), which has been rendered replication-defective for safe laboratory use. The GFP gene, originally isolated from the jellyfish Aequorea victoria, serves as a versatile reporter protein due to its intrinsic fluorescence, allowing real-time visualization of gene expression, cellular localization, and transduction efficiency without the need for additional substrates. The adenoviral backbone efficiently transduces both dividing and non-dividing cells, making it a powerful gene delivery tool.

Human GFP adenoviral particles are widely used in biomedical research for a variety of applications. Primarily, they serve as benchmarks for optimizing gene delivery systems, as GFP expression enables researchers to monitor transduction efficiency and duration in a variety of cell types, including primary cells and difficult-to-transfect cell lines. In functional studies, these particles can be used to label cells for tracking migration, proliferation, or differentiation in vitro and in vivo, such as in tumor xenograft models or stem cell therapy. In addition, they are also valuable in co-transduction experiments, and GFP can be used as a control to normalize data when studying other target genes. In addition to basic research, GFP adenoviral particles are also used in preclinical gene therapy development to evaluate vector tropism, immune response, and safety.

Hepatocyte growth factor (HGF) alleviates acute and chronic inflammation in experimental inflammatory bowel disease, glomerulonephritis, and airway inflammation. Here, researchers evaluated the anti-inflammatory role of HGF in postischemic heart failure. The left anterior descending coronary artery of rats was ligated, and adenovirus containing human HGF (Ad-HGF) or control GFP adenovirus (Ad-GFP) was injected into the myocardium. Four weeks after injection, left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) were significantly increased in Ad-HGF-treated animals compared with the Ad-GFP group. HGF gene therapy improved ventricular geometry, with a significant decrease in left ventricular end-diastolic diameter (LVEDD) and myocardial collagen deposition. After 4 weeks of Ad-HGF treatment, mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β were significantly reduced in the non-infarcted area. Four weeks after infarction, changes in TNF-α, IL-6, and IL-1β levels in the non-infarcted area were positively correlated with LVEDD. Treatment of acute myocardial infarction (AMI) with Ad-HGF in the early stage of myocardial infarction can reduce the levels of proinflammatory cytokines and protect cardiac function. These results suggest that Ad-HGF gene therapy alleviated ventricular remodeling after infarction by reducing inflammation.

Myocardial infarction rats were treated with Ad-GFP and Ad-HGF for 4 weeks. Hearts in the GFP group (Figure 1A) exhibited significant LV dilatation compared to the sham group (Figure 1C), and HGF treatment (Figure 1B) reduced the LV cavity. Histological analysis showed that the infarct area was significantly increased and the thickness of the left ventricular free wall was significantly reduced in the GFP group. HGF significantly reduced the infarct area and increased the wall thickness (Figures 1D and 1E). The histological differences between myocardial infarction rats treated with Ad-HGF and Ad-GFP were further investigated. Histological changes included myocardial fiber fragmentation and a large number of inflammatory cell infiltration (Figures 1F, 1G, and 1H). The number of lymphocytes in the infarct area of the GFP group was significantly increased compared with the sham group. Ad-HGF treatment significantly protected myocardial cells and reduced inflammatory cell infiltration (Figure 1I).

Figure 1. Cardiac morphology and hematoxylin and eosin staining 4 weeks after treatment.Figure 1. Cardiac morphology and hematoxylin and eosin staining 4 weeks after treatment. (Rong S, et al., 2018)

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