Genetic Knockout Comparison
Genetic Knockout Comparison - Related Videos
11:53
A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
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Cited by 17 •
2017
This protocol describes the steps for cloning multiple single guide RNAs into one guide RNA concatemer vector, which is of particular use in creating multi-gene knockouts using CRISPR/Cas9 technology. The generation of double knockouts in intestinal organoids is shown as a possible application of this method.
09:36
Genetic Engineering of Model Organisms
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2023
Transgenesis, or the use of genetic engineering to alter gene expression, is widely used in the field of developmental biology. Scientists use a number of approaches to alter the function of genes to understand their roles in developmental processes. This includes replacement of a gene with a nonfunctional copy, or adding a visualizable tag to a gene that allows the resultant fusion protein to be tracked throughout development. In this video, the viewers will learn about the principles behind...
10:59
Production of Apolipoprotein C-III Knockout Rabbits using Zinc Finger Nucleases
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Cited by 32 •
2013
Recent development in gene targeting tools makes production of knockout (KO) rabbits possible. In the present work, we generated five Apolipoprotein (Apo) C-III KO rabbits using Zinc Finger Nucleases (ZFN). This work demonstrated that ZFN is a highly efficient method to produce KO rabbits.
10:38
An Overview of Genetic Engineering
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2023
Genetic engineering – the process of purposefully altering an organism’s DNA – has been used to create powerful research tools and model organisms, and has also seen many agricultural applications. However, in order to engineer traits to tackle complex agricultural problems such as stress tolerance, or to realize the promise of gene therapy for treating human diseases, further advances in the field are still needed. Important considerations include the safe and efficient delivery of genetic...
12:25
Epigenome-Wide CRISPR-Cas9-Based Knockout Screens on Chemoresistant Cells
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2026
Here, we present a protocol for generating chemoresistant cells using a dose-escalation approach, followed by a CRISPR/Cas9-based screen using a focused sgRNA library targeting epigenetic modifiers to identify regulators of acquired chemoresistance. The protocol also provides multiple optimization points tailored to chemoresistant cell models, offering a robust framework for researchers investigating resistance mechanisms.
FAQs
Constitutive knockouts lack gene function throughout development, whereas conditional knockouts disrupt the gene in selected tissues or under defined conditions. This difference changes how researchers interpret resulting traits because the comparison can distinguish effects associated with complete, lifelong loss from effects observed only in a particular biological setting. The choice therefore depends on the question being investigated.
Controls provide the reference needed to determine whether a cellular, organismal, or biological trait is associated with loss of the selected gene. Researchers compare the knockout model with an unmodified or other control model rather than interpreting the knockout phenotype in isolation. This comparison strengthens the connection between gene disruption and the observed outcome.
A knockout can be generated by disrupting the selected gene through targeted deletion or insertion. CRISPR-based editing is one approach included in this strategy, allowing researchers to create a model in which the gene no longer performs its normal function. Comparing models produced through these disruptions helps connect the altered genetic state with changes in cells, organisms, or traits.
Researchers first select the gene and establish a model in which its function is disrupted through targeted deletion, insertion, or a CRISPR-based approach. They then compare the resulting cells, organisms, or biological traits with unmodified or control models. The final comparison focuses on differences that may reveal how the selected gene contributes to the system under study.
A conditional knockout is useful when the investigation requires gene disruption in a selected tissue or under a defined condition. A constitutive knockout instead removes gene function throughout development. Choosing between them allows the experimental design to match the biological question, particularly when researchers need to examine a location-specific or condition-specific effect rather than a broadly distributed one.
By showing how loss of a selected gene changes cells, organisms, or biological traits, these comparisons can help clarify the gene’s function in a disease-related model. The resulting evidence may also help researchers assess whether the gene represents a potential therapeutic target. The approach therefore connects controlled genetic disruption with disease modeling and treatment-oriented investigation.