Cat.No. : AAV00077Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 1 Storage: -80 ℃
| Cat. No. | AAV00077Z |
| Description | CAG-ChIEF-tdTOMATO AAV (Serotype 1) is the serotype 1 AAV which expresses a hybrid of Channelrhodopsin-1/2 under CAG promoter with tdTOMATO tag. ChIEF is a hybrid of channelrhodopsin 1 and 2 with additional L170I mutation (ChR1 numbering) or at L131I (ChR2-numbering), leading to large currents in oocytes and HEK-cells and almost wild type like open state life time. ChIEF function as light-gated ion channels and very useful for many bioengineering and neuroscience applications such as photostimulation of neurons for probing of neural circuits. Using tdTOMATO tagged ChIEF, light-stimulated axons and synapses can be identified in intact brain tissue. This is useful to study the molecular events during the induction of synaptic plasticity. ChIEF has also been used to map long-range connections from one side of the brain to the other, and to map the spatial location of specific inputs on the dendritic tree of individual neurons. |
| Serotype | AAV Serotype 1 |
| Target Gene | CAG-ChIEF-tdTOMATO |
| Product Type | Adeno-associated virus |
| Titer | Varies lot by lot, typically ≥1x10^12 GC/mL |
| Size | Varies lot by lot, for example, 30 μL, 50 μL, 100 μL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality AAV particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between AAV particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in the production of AAV, especially for applications in animal studies and gene therapy. Effective endotoxin quality control is essential in the development and manufacturing of AAV particles. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced AAV particles to ensure regulatory compliance. |
| Purity | AAV purity is critical for ensuring the safety and efficacy of AAV-based applications.AAV capsids are composed of three main protein components, known as viral proteins: VP1, VP2, and VP3. These proteins play a critical role in the structure and functionality of the AAV capsid. Monitoring the VP1, VP2, and VP3 content in AAV preparations is essential for quality control in AAV production. Our AAV particles are tested for showing three clear bands of VP1, VP2 VP3 by SDS-PAGE. |
| Sterility | The AAV virus samples are inoculated into the cell culture medium for about 5 days to detect bacterial and fungal growth. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of AAV to deliver genetic material into target cells or tissues, and assess gene expression and functional activities. |
| Empty vs. Full Capsids | Based-on our proprietary AAV production and purification technology, Creative Biogene can always offer AAV particles with high ratio of full capsids. If required, we can also assess the ratio for a specifc lot of AAV particles by transmission electron microscopy (TEM) or other methods. |
Human-induced pluripotent stem cells (h-iPSCs)-derived engineered cardiac tissues are immature, limiting their ability to regenerate damaged myocardium and serve as a reliable in vitro model for human disease and drug toxicity studies. Here, researchers designed proof-of-principle experiments to successfully transfect desensitization-resistant chimeric channelrhodopsin proteins into h-iPSCs-derived engineered cardiac tissues, and then subjected the engineered cardiac tissues to optical pacing to accelerate maturation. The researchers transfected h-iPSC engineered cardiac tissues with adeno-associated virus-packaged chimeric channelrhodopsin and then validated optical pacing by whole-cell patch clamp. The engineered cardiac tissues were then subjected to chronic optical pacing in vitro from days 7 to 14 at rates above their intrinsic heart rate. Chronically optically paced resulted in improved electrophysiological properties of the engineered cardiac tissues and subtle changes in the expression of some cardiac-related genes, but no changes in active force production and histology. These results validate the feasibility of a novel chronic optical pacing paradigm to explore non-invasive and scalable strategies for optical pacing-induced maturation of engineered cardiac tissues.
After preliminary trials with several AAV constructs and transfection doses, the researchers transfected differentiated h-iPSC-derived cardiac cells with AAV1/2-CAG-ChIEF-tdTomato virus at an MOI of 500 by adding the virus directly to the cell/matrix mixture at the time of ECT formation. All transfected cells expressed the tdTomato reporter, and this MOI did not alter the percentage of CMs within h-iPSC ECTs, and the mean CM fractions were similar in D14 control (n = 9) and C-OP (n = 10) ECTs. D14C-OP ECTs began beating spontaneously after 3 days in culture, similar to control ECTs (Figure 1).
Figure 1. ECT formation and chronic optical pacing (C-OP) protocol. (Dwenger M, et al., 2019)
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
The CAG-ChIEF-tdTOMATO AAV (Serotype 1) has been instrumental in advancing our neuroscience research. Its high transduction efficiency in neuronal tissue has allowed us to achieve precise gene delivery with minimal off-target effects.
Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.
Our promise to you:
Guaranteed product quality, expert customer support.
24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Copyright © Creative Biogene. All rights reserved.
