Crispr/cas9 Mutagenesis
Crispr/cas9 Mutagenesis - Related Videos
07:13
CRISPR-Cas9-based Mutagenesis in the Entomopathogenic Nematode Steinernema hermaphroditum and the Maintenance of Mutant Lines
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2025
This article demonstrates CRISPR-Cas9-mediated genome engineering in Steinernema hermaphroditum, an entomopathogenic (EPN: insect-parasitic) nematode and an emerging genetic model. The described technology is useful for creating mutants, allowing for the elucidation of gene functions in nematode biology that are relevant to mutualistic and parasitic symbiosis.
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.
11:37
QTL Mapping and CRISPR/Cas9 Editing to Identify a Drug Resistance Gene in Toxoplasma gondii
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Cited by 5 •
2017
Details are presented on how QTL mapping with a whole genome sequence based genetic map can be used to identify a drug resistance gene in Toxoplasma gondii and how this can be verified with the CRISPR/Cas9 system that efficiently edits a genomic target, in this case the drug resistance gene.
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...
10:52
Precise Phage Mutagenesis with NgTET-Assisted CRISPR-Cas Systems
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2025
Here, we present a protocol to reduce DNA modifications in bacteriophages using the NgTET enzyme, enabling efficient and scarless CRISPR-Cas mutagenesis. This method facilitates the genetic engineering of phages for applications in biotechnology and phage therapy.
FAQs
Targeting depends on the guide RNA’s sequence complementarity to the selected DNA region. The guide pairs with that sequence, positioning Cas9 so cleavage occurs at the intended genomic site rather than elsewhere in the design. This pairing establishes which locus is examined in a genetics experiment and supports targeted analysis of gene function.
The PAM acts as an essential neighboring sequence for Cas9 recognition. A complementary target is not sufficient on its own: the DNA site must also lie adjacent to a compatible PAM for cleavage to occur. Consequently, PAM availability helps determine which genomic locations can be selected when planning a mutagenesis experiment.
Mutation outcomes reflect how the cell repairs the Cas9-induced break. Error-prone non-homologous end joining can introduce insertions or deletions, changes that may disrupt gene function and support knockout studies. Homology-directed repair instead can use a designed sequence to produce a specified modification, making repair pathway choice central to experimental goals.
Before editing, a genetics study must specify the locus, the guide RNA sequence, the neighboring PAM, and the desired repair outcome. That planning distinguishes experiments intended to disrupt a gene from those designed to install a particular sequence. It also links the molecular design to the biological question, whether gene function, regulation, or a variant is being examined.
Different edit outcomes support different applications. Insertions or deletions produced through error-prone repair can help create gene knockouts for testing whether a gene contributes to a phenotype. Designed sequence incorporation through homology-directed repair can enable precise sequence modifications. Together, these outcomes allow mutagenesis experiments to connect specific genetic changes with gene function and regulation.
In genetics, the method can be applied to functional screens and to studies of disease-associated variants. Screens use targeted changes across genes or sequences to investigate functional effects, while variant-focused editing helps examine how a particular alteration relates to biology. These research uses also inform the development of potential therapeutic strategies, although such strategies remain development goals rather than established treatments.