Crispr Rhodopsin Knockout
Crispr Rhodopsin Knockout - 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.
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.
04:13
CRISPR Concatemer-Mediated Multiple Gene Knockout: A Technique to Simultaneously Knockout Multiple Genes by Non-Homologous End-Joining Pathway in Mouse Intestinal Cells
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2025
This video describes a gene knockout technique using a CRISPR-concatemer to simultaneously knock out multiple genes in cultured mouse intestinal organoid cells. This method is used to knock out a diseased gene and to elucidate the function of a gene and its paralogues.
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.
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.
FAQs
The guide RNA provides the targeting information that brings Cas9 to a selected rhodopsin DNA sequence. Cas9 then makes a double-strand break. During error-prone repair, insertions or deletions may alter the coding sequence, preventing production of functional rhodopsin protein. This links a defined genomic event to the knockout outcome.
Insertions and deletions matter because they can disrupt the rhodopsin coding sequence rather than merely changing an unrelated region. If the altered sequence no longer supports functional protein production, researchers can relate later retinal phenotypes to rhodopsin loss. The repair outcome therefore determines whether editing yields the intended functional deficiency.
A basic workflow starts with a guide RNA directed to rhodopsin DNA. Cas9 uses that targeting information to create a double-strand break, after which cellular error-prone repair can generate insertions or deletions. Researchers interpret disruption of the coding sequence as the molecular basis for preventing functional rhodopsin production.
The most informative developmental readouts are photoreceptor maturation, retinal organization, and light responses. Examining these dimensions together helps distinguish whether rhodopsin loss is associated with changes in how photoreceptors mature, how retinal tissue is organized, or how the retina responds to light. This connects gene disruption with developmental and functional consequences.
Their value lies in separating consequences of rhodopsin loss from general features of retinal development. By studying deficient models, researchers can ask whether altered photoreceptor maturation, retinal organization, or light responses accompany the loss. This supports direct analysis of gene function within developmental biology studies and retinal research contexts.
Findings from rhodopsin-deficient models can clarify how loss of this photoreceptor protein relates to retinal development and function. That makes the approach relevant to research on inherited retinal disorders, where understanding gene effects can inform investigation of potential genetic therapies. The models therefore connect developmental experiments with disease-oriented research.