Genome Knockout
Genome Knockout - Related Videos
09:56
Improved Genome Editing via Oviductal Nucleic Acids Delivery-based In Vivo Electroporation Technique for Knockout Mice Generation
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2025
The efficiency of the Improved Genome Editing via Oviductal Nucleic Acids Delivery (I-GONAD) method is comparable to traditional microinjection, which requires zygote collection from donor females and transfer to pseudo-pregnant females. This protocol demonstrates its effectiveness by introducing CRISPR/Cas9-induced mutations into the ROSA26 locus on chromosome 6.
08:31
Genome Editing
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2023
A well-established technique for modifying specific sequences in the genome is gene targeting by homologous recombination, but this method can be laborious and only works in certain organisms. Recent advances have led to the development of “genome editing”, which works by inducing double-strand breaks in DNA using engineered nuclease enzymes guided to target genomic sites by either proteins or RNAs that recognize specific sequences. When a cell attempts to repair this damage, mutations can be...
08:31
Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
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Cited by 2 •
2013
Single Virus Genomics (SVG) is a method to isolate and amplify the genomes of single virons. Viral suspensions of a mixed assemblage are sorted using flow cytometry onto a microscope slide with discrete wells containing agarose, thereby capturing the virion and reducing genome shearing during downstream processing. Whole genome amplification is achieved using multiple displacement amplification (MDA) resulting in genomic material that is suitable for sequencing.
06:37
Embryo Microinjection and Knockout Mutant Identification of CRISPR/Cas9 Genome-Edited Helicoverpa Armigera (Hübner)
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Cited by 4 •
2021
Presented here is a protocol of Helicoverpa armigera (Hübner) embryo microinjection and knockout mutant identification created by CRISPR/Cas9 genome editing. Mutant insects enable further research of gene function and interaction among different genes in vivo.
07:37
Performing an In Vitro Genome-Wide CRISPR Knockout Screen in Chimeric Antigen Receptor T Cells
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2025
The article describes a protocol for the application of an in vitro model for exhaustion to complete a genome-wide CRISPR knockout screen in healthy donor chimeric antigen receptor T cells.
FAQs
The targeted DNA break creates a defined site where cellular repair can alter the selected gene. If repair is error-prone, insertions or deletions may appear within the edited sequence. This connects the editing event to a specific genetic change rather than relying only on an observed phenotype, providing a molecular basis for examining the gene’s biological effects.
Insertions or deletions within a coding sequence can disrupt the information needed to produce a functional protein. The consequence is not simply a change in DNA appearance; it can remove the protein activity associated with the gene. Examining the resulting biological changes helps researchers connect protein loss with gene function and significance.
Researchers compare a knockout cell, organism, or model system with an unmodified control and examine differences in phenotype. A change observed after the gene is disabled can indicate that the gene contributes to the affected biological process. This comparison supports studies of gene function while providing a reference for interpreting the edited system’s outcome.
A typical approach selects the gene of interest, introduces a targeted DNA break with a CRISPR-Cas system or another genome-editing method, and relies on cellular repair to alter the sequence. Researchers then examine the resulting model for changes associated with gene disruption. The strategy can be applied in cells, organisms, or other biological model systems.
The choice depends on the biological question being investigated. Knockout cells can support analysis of gene-related effects in a cellular setting, whereas organisms or broader model systems can reveal consequences across biological processes. Using different systems allows researchers to examine gene function, disease mechanisms, developmental pathways, or drug responses at appropriate levels of biological organization.
These experiments can clarify the function and biological significance of selected genes by linking their disruption to measurable phenotypic effects. In biology research, that information supports investigation of disease mechanisms, developmental pathways, and responses to drugs. Knockout findings can also identify genes relevant to therapeutic research, particularly when altered phenotypes suggest biological processes worth further study.