Cas Proteins
Cas Proteins - Related Videos
16:08
Microinjection of CRISPR/Cas9 Protein into Channel Catfish, Ictalurus punctatus, Embryos for Gene Editing
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Cited by 34 •
2018
A simple and efficient microinjection protocol for gene editing in channel catfish embryos using the CRISPR/Cas9 system is presented. In this protocol, guide RNAs and Cas9 protein were microinjected into the yolk of one-cell embryos. This protocol has been validated by knocking out two channel catfish immune-related genes.
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.
04:22
A Technique for Gene Editing in Natural Killer Cells Using CRISPR Cas9
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2025
This video demonstrates a technique for Cas9 ribonucleoprotein-mediated genetic modification of primary natural killer (NK) cells. A Cas9 ribonucleoprotein, consisting of a Cas9 endonuclease bound to a guide RNA (gRNA) formed by base pairing a CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA), is introduced into primary natural killer cells via electroporation. The ribonucleoprotein targets and cleaves the host DNA at the target site, leading to gene knockout via modification of the...
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
Cas proteins locate targets through sequence-specific pairing between a CRISPR RNA and a complementary DNA or RNA sequence. This guide-dependent recognition gives the system selectivity: the RNA supplies the targeting information, while the associated protein carries out the relevant molecular action. Consequently, changing the guide can redirect an engineered system toward a different nucleic-acid sequence.
Their natural immune role is organized around genetic memory of earlier infection. Bacteria and archaea acquire information associated with foreign genetic material, then use CRISPR RNA to support recognition of a matching sequence during a later encounter. This links past exposure to sequence-specific defense and explains why Cas systems are studied as models of adaptive immunity in microbes.
Different Cas activities produce different experimental outcomes. A cutting activity can disrupt a selected genetic sequence, whereas binding without cutting can enable targeting or regulation; modification changes the nucleic-acid target in another way. Detection applications use sequence recognition as the informative event. Selecting among these activities therefore depends on whether the goal is alteration, control, or identification.
The outcome depends on the relationship among the chosen Cas protein, its CRISPR RNA guide, and the complementary nucleic-acid target. Researchers must also define the intended action, such as cutting, binding, modification, or regulation. Matching these components to the biological question helps determine whether the experiment changes a sequence, controls its activity, or detects its presence.
A useful plan identifies the DNA or RNA sequence of interest, the CRISPR RNA needed for complementary recognition, and the molecular outcome being measured. The design should also distinguish whether the system is being used to edit, regulate, detect, or study a target. Clarifying these choices connects the engineered system to a specific biological question and interpretable result.
Cas proteins support gene regulation, nucleic acid detection, and broader biotechnology applications in addition to sequence alteration. Researchers can use these capabilities to investigate gene function and disease mechanisms, while engineered systems also inform potential therapeutic strategies. This range makes Cas proteins valuable both as experimental tools and as models for understanding microbial immune biology.