Cas9 Expression
Cas9 Expression - Related Videos
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.
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...
07:44
CRISPR/Cas9 Technology in Restoring Dystrophin Expression in iPSC-Derived Muscle Progenitors
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Cited by 6 •
2019
Here, we present a Cas9-based exon23 deletion protocol to restore dystrophin expression in iPSC from Dmdmdx mouse-derived skin fibroblasts and directly differentiate iPSCs into myogenic progenitor cells (MPC) using the Tet-on MyoD activation system.
FAQs
The level and control of Cas9 expression influence three linked outcomes: how efficiently a target is edited, how specifically the intended genomic site is recognized, and how well unintended genetic changes are minimized. Consequently, expression is not merely a delivery issue; it is an experimental variable that researchers adjust when designing genome-editing studies.
After Cas9 creates the targeted double-strand break, the cell’s own repair pathways determine what sequence remains. Error-prone repair can resolve the break with unintended sequence changes, whereas supplying a donor template enables precise sequence replacement. Thus, the same nuclease activity can support different genetic outcomes depending on whether templated repair is available.
DNA, RNA, and protein are all described as possible forms for providing Cas9 activity to cells. These formats represent alternative expression or delivery choices within the same editing design, while the guide RNA still supplies target-sequence direction. Selecting a format therefore belongs to construct planning, alongside decisions about expression control and repair outcome.
A basic Cas9-editing workflow begins by selecting a genomic sequence, preparing a Cas9 expression format, and providing a guide RNA with complementary sequence. The nuclease then makes a site-specific double-strand break. Researchers may also supply a donor template when precise replacement is desired, after which cellular repair determines the resulting sequence.
In genetics, Cas9 expression supports functional genomics by allowing researchers to alter selected genomic sites and examine resulting genetic effects. The same framework can be used in disease modeling, where a planned sequence change helps investigate disease-related biology. Potential therapeutic research also depends on controlling expression and limiting unintended changes during experimental evaluation.
These experiments can reveal whether a selected genomic target undergoes the intended change, whether repair produces a precise replacement or an error-prone outcome, and how expression control relates to efficiency, specificity, and unintended genetic changes. Such readouts help connect a designed genetic intervention with its consequences in functional genomics and disease-modeling studies.