Cas9 Complex
Cas9 Complex - Related Videos
06:12
Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex
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Cited by 24 •
2018
Pronuclear (PN) injection of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein-9 nuclease (CRISPR/Cas9) system is a highly efficient method for producing genetically engineered golden Syrian hamsters. Herein, we describe the detailed PN injection protocol for the production of gene knockout hamsters with the CRISPR/Cas9 system.
02:15
Nanoblade-Based Delivery of Nucleic Acid Cargo: A Technique to Deliver Cas9-sgRNA Complex to Target Cells via Virus-Like Particles
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2025
This video demonstrates a technique for nanoblade-based delivery of Cas9-sgRNA ribonucleoprotein complex inside target cells for genome editing. Nanoblades are virus-like particles devoid of the ability to multiply and infect neighboring cells, therefore are very useful for rapid and dose-dependent transportation of bio- and nanomaterials.
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...
FAQs
The protospacer-adjacent motif, or PAM, provides a required sequence context next to the DNA region recognized by the guide RNA. Cas9 uses this neighboring signal while testing potential targets, so guide-RNA complementarity alone is not sufficient for cleavage. In biochemical experiments, the PAM helps determine which DNA sites are eligible for recognition and modification.
Cas9 uses two nuclease domains to cut the two DNA strands in a coordinated manner. The HNH domain cleaves one strand, while the RuvC domain cleaves the other. Their combined activity produces a double-strand break at the selected target, creating the molecular lesion that cellular repair pathways can subsequently process.
Cellular repair of the break can produce different genetic outcomes, depending on how the damaged DNA is processed. Repair may disrupt the targeted gene or support introduction of a designed sequence change. Consequently, the cleavage event is only the initiating step, while repair determines much of the final genome-engineering result.
A typical workflow begins by selecting a DNA sequence that is complementary to a guide RNA and positioned next to a compatible PAM. The assembled complex then recognizes the target, unwinds the DNA, and cleaves both strands. Cellular repair follows and can be evaluated for gene disruption or a planned sequence modification.
Researchers can use targeted cleavage to disrupt a selected gene and examine the resulting biological consequences, supporting gene-function studies. The same strategy can help create models carrying particular genetic changes, which supports disease modeling. These applications connect a defined DNA modification with downstream cellular or organismal phenotypes.
The complex provides a biochemical system for studying programmable DNA recognition, strand cleavage, and repair-dependent genetic outcomes. It also underlies research applications in diagnostics and targeted genetic therapies. Ongoing biochemical work focuses on improving specificity and control, which is important when precise modification is required for research or therapeutic development.