Crispr-cas9 Rnp
Crispr-cas9 Rnp - Related Videos
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...
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.
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.
FAQs
Specificity depends on two linked recognition requirements. The guide RNA must pair with the complementary target sequence, while Cas9 must encounter that sequence next to a compatible protospacer adjacent motif, or PAM. This combination helps position the nuclease at the intended genomic site, making guide design and target selection central to precise bioengineering experiments.
The cellular repair pathway influences the result after Cas9 creates a double-strand break. Nonhomologous end joining repairs the break and can disrupt gene function, whereas homology-directed repair can support a targeted sequence change. Bioengineers therefore interpret the edited cells according to whether the objective is gene disruption or a defined genetic modification.
Because the nuclease is delivered as a preassembled complex, its activity is transient rather than sustained through prolonged nuclease expression. This limits the duration of Cas9 exposure and reduces unwanted genetic exposure. That feature is useful when bioengineers want rapid editing while minimizing continued nuclease activity after the intended DNA modification has occurred.
A typical workflow begins by preassembling Cas9 with a guide RNA designed for a complementary genomic target. The resulting complex is delivered into living cells, where Cas9 makes the programmed double-strand break. Cellular repair then generates either gene disruption or a targeted sequence change, and the resulting cells can be used for downstream bioengineering studies.
The desired outcome guides interpretation of the repair process. If the goal is to impair a gene, nonhomologous end joining provides the relevant disruption outcome. If the goal is to introduce a defined sequence change, homology-directed repair is the pertinent pathway. This distinction helps connect the molecular break to the experimental design and measured result.
CRISPR-Cas9 RNPs support several bioengineering uses, including functional genomics, disease modeling, cell-line development, and therapeutic research. In functional studies, edits can help examine gene roles; in model and cell-line work, they can create modified biological systems. Their rapid, transient delivery also makes them relevant to research focused on controlled genome modification in cells or organisms.