Cat.No. : AD00146Z
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
| Cat. No. | AD00146Z |
| Target Gene | CPT1A |
| Product Type | Adenoviral particle |
| Insert | CPT1A |
| 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. |
| Gene Name | CPT1A carnitine palmitoyltransferase 1A (liver) [ Homo sapiens ] |
| Gene Symbol | CPT1A |
| Synonyms | CPT1A; carnitine palmitoyltransferase 1A (liver); CPT1; carnitine O-palmitoyltransferase 1, liver isoform; CPT1 L; L CPT1; CPT I; CPTI-L; carnitine palmitoyltransferase I, liver; carnitine O-palmitoyltransferase I, liver isoform; CPT1-L; L-CPT1; |
| Gene ID | 1374 |
| UniProt ID | P50416 |
| mRNA Refseq | BC000185 |
| Chromosome Location | 11q13.2 |
| Function | carnitine O-palmitoyltransferase activity; identical protein binding; transferase activity, transferring acyl groups; |
| Pathway | AMPK signaling, organism-specific biosystem; Adipocytokine signaling pathway, organism-specific biosystem; Adipocytokine signaling pathway, conserved biosystem; Circadian Clock, organism-specific biosystem; FOXA2 and FOXA3 transcription factor networks, organism-specific biosystem; Fatty Acid Beta Oxidation, organism-specific biosystem; Fatty acid metabolism, organism-specific biosystem; |
| MIM | 600528 |
During chronic kidney disease (CKD), renal tubular epithelial cells (TECs) display a persistent inflammatory and profibrotic response. Fatty acid oxidation (FAO), the major energy source for TECs, is reduced in renal fibrosis and contributes to its pathogenesis. To determine whether FAO gain-of-function (FAO-GOF) could protect against fibrosis, researchers generated a conditional transgenic mouse model in which the fatty acid shuttle enzyme carnitine palmitoyltransferase 1A (CPT1A) was overexpressed in TECs. Cpt1a-knockin (CPT1A-KI) mice subjected to 3 models of renal fibrosis exhibited reduced expression of fibrotic markers, attenuated proinflammatory responses, and reduced epithelial cell damage and macrophage influx. Protection against fibrosis was also observed when Cpt1a overexpression was induced after FAN. FAO-GOF restored oxidative metabolism and mitochondrial number, enhanced bioenergetics, and increased palmitate oxidation and ATP levels, changes that were also recapitulated in TECs exposed to profibrotic stimuli. Studies in patients have shown reduced CPT1 levels and increased accumulation of short- and medium-chain acylcarnitines, reflecting impaired FAO in human CKD. Therefore, FAO-GOF-based strategies may be a powerful alternative to combat intrinsic fibrosis in CKD.
To confirm the metabolic functional consequences of CPT1A overexpression in a human setting, the researchers examined OCR and ECAR in the human renal tubular epithelial cell line HKC-8. Adenovirus carrying CPT1A (AdCPT1A) or adenovirus control (AdControl) as a negative control was used to infect HKC-8 cells. In HKC-8 cells expressing CPT1A, CPT1A protein levels were 4-fold higher than in control cells (Figure 1A and B). HKC-8 cells transduced with CPT1A exhibited reduced TGF-β1-induced FAO inhibition (Figure 1C) and inhibition of glycolysis, which was reflected in ECAR levels (Figure 1D). As expected, FAO rates were 2-fold higher in HKC-8 cells expressing AdCPT1A, which was confirmed by measuring 14C-palmitate-derived 14CO2 (Figure 1E). Consistently, CPT1A overexpression increased the fluorescence resonance energy transfer (FRET) signal of a specific ATP sensor and promoted the reduced decay in the presence of TGF-β1 .
Figure 1. HKC-8 cells transduced with CPT1A exhibit reduced TGF-β1-induced FAO inhibition and fibrogenic transformation. (Miguel V, et al., 2021)
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Using these adenoviral particles, we achieved robust CPT1A overexpression with minimal off-target effects. Great product for metabolic research!
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