Knockout Mutants
Knockout Mutants - Related Videos
08:25
Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
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Cited by 6 •
2017
The genotyping technique described here, which couples fluorescent polymerase chain reaction (PCR) to capillary gel electrophoresis, allows for high-throughput genotyping of nuclease-mediated knockout clones. It circumvents limitations faced by other genotyping techniques and is more cost effective than sequencing methods.
11:45
Generation of Marked and Markerless Mutants in Model Cyanobacterial Species
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Cited by 36 •
2016
Introducing multiple genomic alterations into cyanobacteria is an essential tool in the development of strains for industrial and basic research purposes. We describe a system for generating unmarked mutants in the model cyanobacterial species Synechocystis sp. PCC6803 and marked mutants in Synechococcus sp. PCC7002.
09:17
Generation of Maternal Mutants Using zpc:cas9 Knock-in Zebrafish
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2025
Here, we describe a protocol for generating maternal mutant that couples a stable zpc:cas9 knock-in line with Tol2-mediated delivery of sgRNA expression cassettes.
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.
11:35
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Cited by 6 •
2017
Recent advances in the ability to genetically manipulate somatic cell lines hold great potential for basic and applied research. Here, we present two approaches for CRISPR/Cas9 generated knockout production and screening in mammalian cell lines, with and without the use of selectable markers.
FAQs
The key inference comes from comparing the altered system with an otherwise similar control. If the two differ in growth, development, physiology, or a disease-related trait, the difference links those outcomes to the missing gene activity. This comparison turns a genetic change into functional evidence and helps distinguish the gene’s contribution from background biological variation.
Removing or altering a gene’s coding sequence is important because the change is designed to produce loss of gene activity. Researchers can then examine consequences associated with that loss rather than merely observing a naturally occurring difference. The resulting comparison helps connect a specific genetic alteration with cellular or organismal traits.
Phenotypes provide the observable link between gene activity and biological pathways. Changes in growth or development can indicate roles in broader organismal processes, whereas physiological or disease-related changes can point to effects at the cellular or systems level. Examining these outcomes across a knockout mutant and its control supports genetic pathway analysis.
A typical study begins by selecting a gene, introducing a targeted mutation, and obtaining the resulting organism, cell, or tissue for comparison. Researchers then assess relevant traits against an otherwise similar control, focusing on growth, development, physiology, or disease-related characteristics. The observed differences are interpreted as evidence about the gene’s biological contribution.
Knockout mutants are especially useful in functional genomics, where researchers connect genes with their biological roles. By examining what changes after gene activity is lost, investigators can study relationships among genes and investigate pathways rather than treating gene sequences as isolated information. This makes the approach valuable for systematic analysis of biological function.
In disease modeling, researchers examine whether loss of a gene produces disease-related traits or other relevant physiological changes. These models can clarify how the gene contributes to disease processes and support evaluation of potential drug targets. Their value lies in linking a defined genetic alteration to measurable biology relevant to therapeutic investigation.