Adenovirus-mediated Gene Deletion
Adenovirus-mediated Gene Deletion - Related Videos
09:16
Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells
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Cited by 2 •
2022
This protocol describes the process of the generation and characterization of mouse urothelial organoids harboring deletions in genes of interest. The methods include harvesting mouse urothelial cells, ex vivo transduction with adenovirus driving Cre expression with a CMV promoter, and in vitro as well as in vivo characterization.
11:24
Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs
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Cited by 8 •
2017
Gene deletion mutants generated through homologous recombination are the gold standard for gene function studies. The OSCAR (One Step Construction of Agrobacterium-Recombination-ready-plasmids) method for rapid generation of deletion constructs is described. Agrobacterium mediated fungal transformation follows. Finally, a PCR based confirmation method of gene deletions in fungal transformants is presented.
03:36
Generating Gene Deletions in E. coli via Phage-Mediated Transfer of an Excisable Antibiotic Cassette
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2026
Source: Saragliadis, A., et al. Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes. J. Vis. Exp. (2018)This video demonstrates the phage-mediated transfer of donor DNA carrying an excisable antibiotic resistance cassette into Escherichia coli. The integrated cassette replaces the target gene via homologous recombination, resulting in a stable gene deletion.
11:00
Adenovirus-mediated Genetic Removal of Signaling Molecules in Cultured Primary Mouse Embryonic Fibroblasts
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Cited by 5 •
2010
In this video we use an adenovirus carrying the Cre recombinase gene to infect primary mouse embryonic fibroblasts carrying a floxed Rac1 allele.
11:13
Dissection of Adult Mouse Utricle and Adenovirus-mediated Supporting-cell Infection
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Cited by 18 •
2012
Mechanosensory hair cells are the receptor cells of the inner ear. The best-characterized in vitro model system of mature mammalian hair cells utilizes organ cultures of utricles from adult mice. We present the dissection of the adult mouse utricle, and we demonstrate adenovirus-mediated infection of supporting cells in cultured utricles.
FAQs
Cre recombinase recognizes two loxP sites positioned around the target gene. After the adenoviral vector delivers Cre into infected cells, the enzyme acts on those sites and excises the DNA between them. This removes the selected genetic segment from affected cells, allowing researchers to connect loss of that gene with subsequent cellular changes.
The loxP sites define the DNA segment that Cre recombinase can remove. When they flank the target gene, Cre has the necessary boundaries for excision; without that arrangement, the intended conditional deletion strategy cannot selectively remove the intervening sequence. Their placement therefore determines which gene is disrupted when cells receive the adenoviral machinery.
The outcome depends on which cells receive the adenoviral vector and when delivery occurs. Restricting infection to selected cells can reveal gene functions in particular tissues, while choosing the time of delivery can separate gene disruption from earlier developmental events. These features help researchers examine effects that a permanent, organism-wide deletion might obscure.
A typical workflow begins with placing loxP sites around the gene of interest, followed by delivering a Cre-expressing adenoviral vector to the selected cells. Infection introduces the recombinase, which excises the intervening target DNA. Researchers then examine the treated cells for responses associated with loss of the gene, using the chosen timing and cellular context.
This strategy is useful when a permanent germline deletion is impractical or produces developmental effects that complicate interpretation. Researchers can disrupt the gene in selected cells and at a chosen time instead of removing it throughout development. That flexibility supports experiments focused on later gene function, tissue-specific biology, or responses that emerge after targeted disruption.
Adenovirus-mediated gene deletion can support studies of gene function, disease mechanisms, and cellular responses to targeted gene loss. By examining selected cells after Cre-mediated excision, investigators can assess consequences in a defined biological setting rather than relying only on a permanent germline model. The approach is therefore suited to linking gene disruption with tissue or cell behavior.