Crispr Gene Targeting
Crispr Gene Targeting - Related Videos
07:27
CRISPR Interference-Based Gene Silencing: A Technique for Targeted Repression of Gene Function in Pathogenic Leptospira
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2025
This video demonstrates a CRISPR interference-based gene silencing in pathogenic Leptospira. Targeted repression of gene expression using a CRISPR/Cas system that blocks mRNA formation helps identify candidate genes for pathological and pharmacological studies.
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...
08:56
Recombineering and Gene Targeting
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2023
One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based...
02:58
CRISPR-Mediated Base Editing Tools: A Genome Editing Technique to Induce Targeted Base Substitution
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2025
This video explains the concept of CRISPR-mediated cytosine base editors for inducing targeted nucleotide substitution.
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.
FAQs
The guide RNA carries a sequence complementary to the selected DNA target and positions the CRISPR-associated nuclease at that site. Targeting also depends on a nearby protospacer-adjacent motif, or PAM, which the nuclease recognizes before cutting. Together, guide complementarity and PAM availability constrain which genomic sequences can be modified and where the DNA break occurs.
After the nuclease cuts the target, the cell can repair the break through error-prone nonhomologous end joining or template-directed homology-directed repair. Nonhomologous end joining can disrupt the targeted gene through repair-associated sequence changes, whereas homology-directed repair can support a defined sequence change when a repair template guides restoration. The pathway therefore influences whether editing produces disruption or precision modification.
A targeted DNA break converts a chosen genomic site into a testable perturbation. If repair disrupts the gene, researchers can examine how loss of its function affects gene regulation, cell fate, or tissue formation. If repair introduces a defined change or label, the same locus can be studied for regulatory activity and developmental behavior in relevant biological systems.
By modifying selected genes or regulatory elements, researchers can connect genomic sequences with developmental outcomes. Disruption can reveal whether a gene is required, while defined sequence changes or labels can help investigate regulatory elements. Embryos, stem cells, and model organisms provide contexts in which these modifications can be related to cell fate decisions, tissue formation, and developmental disorders.
A typical workflow begins by selecting a developmental gene or regulatory element and identifying a compatible target sequence near a PAM. Researchers then direct a CRISPR-associated nuclease to that site, allow cellular repair to generate either disruptive or defined changes, and examine the resulting developmental phenotype or regulatory behavior in embryos, stem cells, or model organisms.
The choice depends on the biological question. Gene disruption is useful for testing whether a gene is required for a developmental process. A defined sequence change can examine the effect of a specific alteration, while labeling a regulatory element supports investigation of its activity or behavior. These outcomes allow separate analysis of gene function, sequence requirements, and developmental regulation.
Edited developmental systems can reveal how particular genes or regulatory elements influence gene regulation, cell fate, and tissue formation. Comparing disrupted, precisely altered, or labeled loci can distinguish functional loss from sequence-specific effects and regulatory activity. Such experiments can also connect genomic changes with developmental disorders, especially when the modified system reproduces relevant developmental processes.