Hypoxia-sensitive Car
Hypoxia-sensitive Car - Related Videos
09:12
Generation and Functional Verification of Hypoxia-Sensitive Chimeric Antigen Receptor-T Cells
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Cited by 1 •
2024
Here we present a protocol for the generation and functional verification of hypoxia-sensitive chimeric antigen receptor (CAR)-T cells. This protocol presents the lentivirus-based generation of hypoxia-sensitive CAR-T cells and their characterization, including the validation of hypoxia-dependent CAR expression and selective cytotoxicity.
07:01
Induction and Testing of Hypoxia in Cell Culture
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Cited by 247 •
2011
Here we propose simple methods to induce hypoxia in cell cultures and simple tests to evaluate the hypoxic status of the cultures.
02:55
Genetically Modifying CAR T Cells Using a CRISPR-Cas9 System
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2025
The video outlines a process for creating genetically modified CAR T cells through the CRISPR-Cas9 System. Infecting T cells with CRISPR and CAR lentiviruses results in modifications to the target gene and the synthesis of a chimeric antigen receptor or CAR, ultimately leading to the formation of genetically modified CAR T cells.
03:32
A Technique for the Generation of Antigen-Specific CAR T Cells
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2025
This video demonstrates a technique for generating antigen-specific chimeric antigen receptor (CAR) T cells through retroviral transduction. Activated murine CD8+ T cells are added to a microwell plate coated with recombinant fibronectin fragments. Retroviral vectors containing transgenic RNA encoding an antigen-specific chimeric antigen receptor (CAR) are then introduced, and the plate is centrifuged to facilitate transduction. After incubation, the transduced cells express antigen-specific...
08:01
Induction of Hypoxia in Living Frog and Zebrafish Embryos
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Cited by 5 •
2017
We introduce a novel hypoxic chamber system for use with aquatic organisms such as frog and zebrafish embryos. Our system is simple, robust, cost-effective and allows the induction and sustainment of hypoxia in vivo and for up to 48 h. We present 2 reproducible methods to monitor the effectiveness of hypoxia.
FAQs
Hypoxia-responsive elements can be used to control transcription of the CAR gene. Their placement makes receptor production responsive to local oxygen conditions rather than independent of the surrounding environment. In cancer research, this provides a transcriptional layer of control that links CAR expression to oxygen-poor tumor regions and helps investigators examine microenvironment-dependent immune-cell activity.
An oxygen-dependent degradation domain provides an additional level of receptor control by promoting CAR breakdown under normoxia and stabilizing the receptor when oxygen is scarce. This changes receptor availability after the protein is produced, complementing transcriptional regulation. Consequently, oxygen conditions can influence both CAR expression and the persistence of the receptor on immune cells.
Using both mechanisms connects oxygen conditions to two stages of receptor regulation: CAR production through hypoxia-responsive elements and receptor stability through an oxygen-dependent degradation domain. This layered design may provide more environmental control than relying on either mechanism alone. In tumor studies, it helps examine how local oxygen availability shapes immune-cell recognition and cytotoxicity.
A hypoxia-sensitive CAR is intended to make immune-cell activity conditional on the tumor microenvironment, whereas a continuously active receptor is not linked to oxygen status in the same way. The key distinction is environmental regulation: oxygen-poor regions can favor receptor availability or expression, while well-oxygenated conditions can reduce it through the engineered controls.
Researchers can introduce oxygen-responsive control at the transcriptional level, the receptor-stability level, or both. Hypoxia-responsive elements regulate CAR transcription, while oxygen-dependent degradation domains regulate receptor breakdown according to oxygen availability. This design workflow allows investigators to connect receptor presence with local conditions before studying how engineered immune cells behave in tumor and well-oxygenated settings.
Oxygen-poor regions are a tumor-microenvironment condition that this receptor design is intended to address functionally. Linking CAR availability to hypoxia may focus immune-cell recognition and cytotoxicity where oxygen is scarce, while reducing activity in healthy, well-oxygenated tissues. The approach therefore seeks greater tumor selectivity by using local oxygen conditions as a regulatory signal.
It provides a platform for examining how oxygen conditions influence cellular immunotherapies. By relating hypoxia to CAR transcription, receptor breakdown, and immune-cell activity, researchers can study whether local microenvironmental differences alter recognition and cytotoxicity. The design is therefore useful not only as a therapeutic strategy, but also as a model for testing microenvironment-dependent control.