Cat.No. : LV00946Z
Titer: ≥1*10^7 TU/mL / ≥1*10^8 TU/mL / ≥1*10^9 TU/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | LV00946Z |
| Description | This lentivirus contains no insert gene under the control of CMV promoter and can be used for negative control. |
| Target Gene | Empty |
| Titer | Varies lot by lot, for example, ≥1*10^7 TU/mL, ≥1*10^8 TU/mL, ≥1*10^9 TU/mL etc. |
| Size | Varies lot by lot, for example, 100 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 lentivirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between lentivirus particle lots. | |
| Mycoplasma | Creative Biogene routinely tests for mycoplasma contamination using a mycoplasma detection kit. Cell lines are maintained for approximately 20 passages before being discarded and replaced with a new vial of early passage cells. Approximately 2 weeks after thawing, cell culture supernatants are tested for mycoplasma contamination. Creative Biogene ensures that lentiviral products are free of mycoplasma contamination. |
| Purity | Creative Biogene evaluates the level of impurities, such as residual host cell DNA or proteins, in prepared lentiviral vectors to ensure they meet quality standards. |
| Sterility | The lentiviral samples were inoculated into cell culture medium for about 5 days and the growth of bacteria and fungi was tested. Creative Biogene ensures that the lentiviral products are free of microbial contamination. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of lentivirus to deliver genetic material into target cells, and assess gene expression and functional activities. |
| Proviral Identity Confirmation | All Creative Biogene lentiviral vectors are confirmed to have correctly integrated provirus using PCR. This test involves transducing cells with serial dilutions of the lentiviral vector, harvesting the cells a few days later, and isolating genomic DNA. This DNA is then used as a template to amplify a portion of the expected lentiviral insert. |
The risk of developing primary open-angle glaucoma (POAG) is associated with the degree of ocular hypertension (OHT) and the concentration of transforming growth factor-β2 (TGFβ2) in the aqueous humor. Here, molecular, optical, electrophysiological, and tonometry strategies were used to determine the role of TGFβ2 in transcriptional and functional expression of mechanosensitive channel isoforms, while studying trabecular meshwork (TM) contractility in a biomimetic hydrogel, and intraocular pressure (IOP) regulation in a TGFβ2-induced OHT mouse model. TGFβ2 upregulated the expression of TRPV4 and PIEZO1 transcripts and enhanced functional TRPV4 activation over time. TRPV4 activation induced TM contractility, while pharmacological inhibition suppressed TGFβ2-induced hypercontractility and abolished OHT in eyes overexpressing TGFβ2. Trpv4-deficient mice were resistant to TGFβ2-driven IOP elevation. Nocturnal OHT had no effect on TGFβ-induced OHT. These studies establish a fundamental role for TGFβ as a mechanosensing regulator in non-excitable cells, identify TRPV4 channels as the ultimate common mechanism underlying TM contractility as well as circadian and pathological OHT, and provide insights into future therapeutic approaches that could reduce IOP in the large population of hypertensive glaucoma patients who are resistant to current treatments.
To test whether TRPV4 contributes to TGFβ2-induced ocular hypertension (OHT) in vivo, the researchers utilized a lentiviral TGFβ2 overexpression model developed by Patil et al. Adult C57BL/6J mice were intravitreally injected with lentivirus overexpressing constitutively active human TGFβ2 (LV-TGFβ2). One week after transduction, IOP was significantly increased in eyes injected with LV-TGFβ2, but not in contralateral eyes injected with lentivirus containing a scrambled transgene (LV-CMV-Null-Puro, LV-Ctrl) (Figure 1A). At 2 weeks after transfection, IOP in LV-TGFβ2 eyes reached 19.9 ± 4.7 mmHg, whereas IOP in LV-Control eyes remained at control levels (14.0 ± 1.2 mmHg), ΔTGF-Ctrl = 5.9 mmHg . The intraocular pressure remained elevated for 4 weeks after the injection. HC-06 (100 µM) microinjection into the anterior chamber of LV-TGFβ2 and LV-Ctrl eyes lowered IOP in LV-TGFβ2 eyes to 12.2 ± 1.7 mm Hg after 24 hours with no difference observed in IOP from LV-Ctrl eyes. LV-Ctrl eyes remained close to pre-injection levels after HC-06 treatment ( Figure 1A-B). IOP in LV-TGFβ2 eyes returned to hypertensive levels 1 week after HC-06 injection. These data suggest that selective pharmacological inhibition of TRPV4 can effectively and reversibly block TGFβ2-induced OHT.
Figure 1. TRPV4 activation is necessary to maintain LV-TGFβ2-induced ocular hypertension. (Rudzitis C N, et al., 2024)
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The CMV-Null-Puro Lentivirus from Creative Biogene was instrumental in our gene expression studies. Its high efficiency in transduction allowed us to achieve consistent and reproducible results.
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