Cat.No. : AD00272Z
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. | AD00272Z |
| Target Gene | TWIST1 |
| Species | Human |
| Product Type | Adenoviral particle |
| Insert | TWIST1 |
| 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 | TWIST1 twist homolog 1 (Drosophila) [ Homo sapiens ] |
| Gene Symbol | TWIST1 |
| Synonyms | SCS; ACS3; CRS1; BPES2; BPES3; TWIST; bHLHa38 |
| Gene Description | twist homolog 1 (acrocephalosyndactyly 3; Saethre-Chotzen syndrome) (Drosophila) |
| Gene ID | 7291 |
| UniProt ID | Q15672 |
| mRNA Refseq | NM_000474.3 |
| Protein Refseq | NP_000465.1 |
| Chromosome Location | 7p21.2 |
| Function | E-box binding; bHLH transcription factor binding; protein binding; protein domain specific binding; protein heterodimerization activity; protein homodimerization activity; sequence-specific DNA binding RNA polymerase II transcription factor activity; transcription factor binding; |
| Pathway | Adipogenesis, organism-specific biosystem; HIF-2-alpha transcription factor network, organism-specific biosystem; Neural Crest Differentiation, organism-specific biosystem; Notch-mediated HES/HEY network, organism-specific biosystem; |
| MIM | 601622 |
Dysregulated chondrocyte activation and de-differentiation in articular cartilage are important factors in the pathogenesis of osteoarthritis (OA). Twist-related protein 1 (TWIST1), known as class A basic helix-loop-helix protein 38 (bHLHa38), is required for embryonic development through epithelial-mesenchymal transition (EMT) and regulates mesoderm formation and morphogenesis. Here, researchers show that TWIST1 expression is increased in human OA knee cartilage compared with normal knee cartilage. TWIST1 induces matrix metalloproteinase 3 (MMP3) expression without directly binding to the MMP3 promoter and increases 5-hydroxymethylcytosine (5hmC) levels at the MMP3 promoter. The effect of TWIST1 on the expression of the TET family (TET1, 2, and 3) was measured in TC28 cells stably transfected with TWIST1 and found that TET1 expression was upregulated. TWIST1-dependent upregulation of Mmp3 expression was suppressed in Tet triple KO fibroblasts derived from mouse ES cells. Increased TWIST1 expression is a characteristic of OA-affected cartilage. Thus, the researchers discovered a novel mechanism of catabolic reaction where TWIST1 up-regulates MMP3 expression by enriching 5hmC levels at the MMP3 promoter via TET1 induction. These findings suggest that TWIST1 is an important factor in regulating OA-related gene expression.
Here, researchers confirmed the enrichment of 5hmC in human OA cartilage by immunohistochemistry (Figure 1A). In TC28 cells, bisulfite DNA sequencing analysis showed that MMP3 promoter CpG loci were methylated, and TWIST1-adenovirus (Ad-TWIST1) did not affect the methylation status compared with untreated cells (NTC) and eGFP-adenovirus (Ad-GFP) treated groups (Figure 1B). However, the 5hmC status level around the CCGG site in the MMP3 promoter was specifically increased, but Ad-TWIST1 did not change the 5mC status level compared with Ad-GFP (Figure 1C). In human chondrocytes, TET1 and TET3 mRNA levels increased by approximately 2-fold due to Ad-TWIST1 infection (Figure 1D). These results suggest that TWIST1 promotes 5hmC at specific MMP3 promoter sites through TET1 and TET3 upregulation.
Figure 1. DNA methylation and 5hmC status in the MMP3 promoter. (Hasei J, et al., 2017)
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The adenoviral particles delivered TWIST1 seamlessly into our target cells. High transduction efficiency with minimal optimization needed. Highly recommend!
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