Rosa26-lsl-cas9-egfp Mouse
Rosa26-lsl-cas9-egfp Mouse - Related Videos
04:40
Modeling Prostate Cancer in Genetically-engineered Mouse Models: A CRISPR/Cas9-mediated Localized Gene Editing Technique in Mouse Anterior Prostate Lobe Cells
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2023
This video describes a method for editing specific genes in prostate gland cells using an adenovirus-based delivery system. The approach allows for orthotopic and localized alteration of gene expression utilizing CRISPR technology to develop novel mouse models for prostate cancer.
02:30
A CRISPR-Cas9 Technique for Gene Editing in T Cells
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2025
This video demonstrates an assay for performing gene editing in human T cells using the CRISPR-Cas9 technology. A mixture of primary CD4+ and CD8+ T cells is combined with a CRISPR-Cas9 ribonucleoprotein complex, targeting specific genes for knockout. Upon electroporation, the sgRNA guides Cas9 to the target DNA sequence, creating precise cuts. These cuts are then repaired by the cell's non-homologous end-joining mechanism, leading to gene knockout.
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.
10:07
A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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Cited by 5 •
2017
This protocol outlines the workflow of a CRISPR/Cas9-based gene editing system for the repair of point mutations in mammalian cells. Here, we use a combinatorial approach to gene editing with a detailed follow-on experimental strategy for measuring indel formation at the target site—in essence, analyzing onsite mutagenesis.
02:36
Establishing a Murine Tumor Cell Line In Vitro Using a Virus-Cas9 System
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2026
Source: Prasad, M. et al. In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated Virus-Cas9 System. J. Vis. Exp. (2020)This video demonstrates the in vitro transformation of murine epithelial cells through viral delivery of Cre recombinase and mutation-inducing components. It outlines the steps for Cas9 activation, cancer-gene modification, and the establishment of a tumorigenic murine cell line.
FAQs
Cre recombinase removes the loxP-flanked transcriptional stop cassette positioned before Cas9 and EGFP at the Rosa26 locus. This event permits expression only in cells where Cre is active, creating a route to restrict genome editing and fluorescent labeling to selected tissues or cell populations. The resulting spatial control is central to modeling cancers that arise from defined somatic compartments.
Cas9 provides the genome-editing activity, while EGFP marks cells that have undergone activation of the cassette. Their coordinated expression links the capacity for guide RNA-directed DNA cleavage with a visible label for the edited population. This pairing helps researchers track those cells as tumors initiate and progress, rather than relying only on later genetic analysis.
Guide RNAs direct Cas9 to chosen DNA sequences, so their targets determine which genes can be disrupted or altered in activated somatic cells. Researchers can therefore design editing strategies around tumor-suppressor loss or oncogene activation. The targeted cleavage connects a specific genetic change with subsequent observations of tumor biology, genetic drivers, and disease progression.
A study first establishes which tissue or cell population should activate the system through Cre, then introduces or uses guide RNA targeting the gene of interest. Activated cells express Cas9 and EGFP, allowing targeted editing and identification of the affected population. Researchers can subsequently follow tumor initiation, progression, and responses to experimental treatments in that context.
The model is suited to testing how defined genetic changes influence tumor initiation and progression in selected tissues. It can support investigations of tumor-suppressor loss, oncogene activation, genetic drivers, and the behavior of edited cell populations over time. Because editing occurs in somatic cells, researchers can examine cancer development within tissue-specific biological settings.
EGFP labeling provides a way to identify and track cells that activated Cas9, enabling researchers to follow their contribution during tumor development. This supports analysis of how edited populations change as tumors progress and how they respond to therapeutic interventions. The system therefore connects a defined genetic manipulation with observable population behavior in cancer research.