Vivo V9 4gb
Esam Rosado
The In Vivo Therapeutics (IVT) team maintains and distributes on-site breeding colonies of specialized immunocompetent (BoyJ, C57BL/6, BoyJ/C57 F1 cross) and immunodeficient (NSG and NRG) mouse strains.
As requested and coordinated by research programs, we support the set up and validation of syngeneic mouse tumor models and development of patient-derived human xenografts.
We manage all aspects of the IVIS SpectrumCT optical imaging system to evaluate tumor response in mouse models via bioluminescence and fluorescence modalities.
The IVT team also manages the radiation facility and performs irradiation services needed for in vitro and in vivo experiments, including hematopoietic stem-cell transplantations.
For more information, contact IVT manager Tony Sinn at als...@iu.edu or 317-274-8811.
The core is funded, in part, by grants and contracts of the investigators who utilize the facility. Due to the variety of in vivo models and drug regimens requested, the specific costs of a study will be calculated once the study design is finalized.
NanoLuc luciferase provides a new reporter option for advanced in vivo imaging applications. Its small size is ideal for engineering into compact genomes, and ATP-independence allows in vivo monitoring of both intracellular and extracellular events. Substrate specificity makes NanoLuc luciferase an ideal complementary reporter to firefly luciferase for an improved dual-luciferase in vivo imaging solution.
The Nano-Glo Fluorofurimazine In Vivo Substrate (FFz) is an optimized reagent designed specifically for in vivo detection of NanoLuc luciferase, NanoLuc fusion proteins or reconstituted NanoBiT luciferase. This optimized, aqueous-soluble, in vivo detection reagent provides increased substrate bioavailability for bright, stable signals and offers flexible delivery options with handling requirements compatible with in vivo workflows.
The Nano-Glo Fluorofurimazine In Vivo Substrate can be delivered via multiple routes of administration. Each method offers different signal intensity and kinetics, allowing flexibility between experimental systems.
NanoLuc signal of 4T1 primary tumors in BALB/c mice. Cells expressing NanoLuc luciferase were orthotopically implanted into the #3 mammary fat pad of female BALB/c mice, and tumors were allowed to grow for 22 days. Mice were injected with FFz via i.p. (top image; circles) or i.v. injection (bottom image; squares). Bioluminescence imaging was done at the University of Wisconsin Small Animal Imaging and Radiotherapy Facility.
In vivo whole-body imaging of mice injected with AAV9-NanoLuc-HaloTag viral particles. Mice were injected with increasing levels of AAV9-NanoLuc-HaloTag viral particles. Nano-Glo FFz (0.44μmol) was injected 7- and 13-days post-transduction followed by in vivo imaging. Tissue tropism starts to appear at 7 days, and a stronger signal and more extensive tissue tropism is visible after 13 days. Bioluminescence imaging was done at the University of Wisconsin Small Animal Imaging and Radiotherapy Facility.
NanoLuc reporters can be multiplexed with firefly reporters to enable two-population bioluminescent imaging in animals. This publication used a NanoLuc reporter to track tumor size and a firefly substrate to visualize CAR-T cells in the same animal over multiple days:
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For uses of Nano-Glo-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
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RNA and DNA expression vectors containing genes for chloramphenicol acetyltransferase, luciferase, and beta-galactosidase were separately injected into mouse skeletal muscle in vivo. Protein expression was readily detected in all cases, and no special delivery system was required for these effects. The extent of expression from both the RNA and DNA constructs was comparable to that obtained from fibroblasts transfected in vitro under optimal conditions. In situ cytochemical staining for beta-galactosidase activity was localized to muscle cells following injection of the beta-galactosidase DNA vector. After injection of the DNA luciferase expression vector, luciferase activity was present in the muscle for at least 2 months.
CHOP has been at the forefront of major in vivo gene therapy milestones over the past two decades. To date, we have treated more than 120 patients from 15 countries with in vivo gene therapies. Led by experts with more than 10 years of gene therapy clinical trial experience, the CIGT team aims to offer more breakthrough therapies to our patients sooner, by streamlining the startup of complex gene therapy trials and ensuring licensed gene therapies are available in our clinics.
Led by experts with more than 10 years of gene therapy clinical trial experience, the CIGT team aims to offer more breakthrough therapies to our patients sooner, by streamlining the startup of complex gene therapy trials and ensuring licensed gene therapies are available in our clinics.
National Institute on Deafness and Other Communication Disorders (NIDCD) Funding Opportunity Title In Vivo High-Resolution Imaging for Inner Ear Visualization (R01 Clinical Trial optional) Activity Code R01 Research Project Grant
This funding opportunity aims to support the development of in vivo high-resolution structural and functional imaging technologies for the living human inner ear. Proposed projects should focus on improving the resolution of current imaging techniques or developing new imaging techniques that can visualize inner ear structures in vivo with significantly greater detail and accuracy than currently possible. Both structural and functional aspects, including visualizing dynamic elements are important to the development of new and improved techniques. Projects may also focus on developing new imaging probes or contrast agents that can enhance visualization of the inner ear structures. Research supported in response to this funding opportunity is expected to significantly advance the ability to visualize auditory and vestibular components, such as hair cells, otoliths, membranes, ions, and vasculature, in detail in awake patients in a clinical setting using non-invasive techniques. To achieve this goal, a multidisciplinary team approach that takes advantage of the expertise of each team member is highly encouraged. Studies in humans and intermediate studies in mammalian animal models may be proposed to develop or advance the needed technology. Any intermediate studies must articulate a clear path of the proposed methodology to application in awake humans or define the limitations and the usefulness in anesthetized humans.
This notice of funding opportunity announcement (NOFO) also supports investigator initiated low-risk clinical trials addressing the development or improvement of human in vivo structural and functional imaging technologies. Clinical trials must meet ALL the following criteria: meet the budget limits of this NOFO, not require FDA oversight, not be an NIH-defined Phase III Clinical Trial, have low risks to subjects, and intend to gather scientific data/evidence to inform subsequent studies. This NOFO also supports low-risk trials determined to be Basic Experimental Studies with Humans (BESH). These studies fall within the NIH definition of a clinical trial and meet the definition of basic research. It is advisable that only one clinical trial be proposed in each application. High-risk clinical trials not meeting all the criteria above are referred to the companion U01 Cooperative Agreement for In Vivo High-Resolution Imaging for Inner Ear Visualization (Clinical Trial required) NOFO RFA-DC-24-005.
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