Crispr-dcas9 Complex
Crispr-dcas9 Complex - Related Videos
06:12
Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex
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Cited by 24 •
2018
Pronuclear (PN) injection of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein-9 nuclease (CRISPR/Cas9) system is a highly efficient method for producing genetically engineered golden Syrian hamsters. Herein, we describe the detailed PN injection protocol for the production of gene knockout hamsters with the CRISPR/Cas9 system.
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.
02:53
CRISPR and crRNAs
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2020
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea. The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
11:53
Substrate Generation for Endonucleases of CRISPR/Cas Systems
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Cited by 4 •
2012
CRISPR/Cas systems mediate adaptive immunity in Bacteria and Archaea. Many Cas proteins are proposed to act as endoribonucleases acting on crRNA precursors of varying length. Here we illustrate three different approaches to generate pre-crRNA substrates for the biochemical analysis of Cas endonuclease activity.
02:55
Genetically Modifying CAR T Cells Using a CRISPR-Cas9 System
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2025
The video outlines a process for creating genetically modified CAR T cells through the CRISPR-Cas9 System. Infecting T cells with CRISPR and CAR lentiviruses results in modifications to the target gene and the synthesis of a chimeric antigen receptor or CAR, ultimately leading to the formation of genetically modified CAR T cells.
FAQs
Targeting is specified by the guide RNA sequence. A region complementary to the chosen DNA sequence enables RNA-DNA base pairing, which positions dCas9 at the intended genomic locus. Once bound, the complex can influence that region, so guide selection determines where transcriptional regulation occurs. This makes guide choice central to the specificity of the regulatory outcome.
Cas9’s catalytic inactivity changes the consequence of target recognition. The protein can bind a selected genomic sequence, but it does not create the DNA cut associated with an active nuclease. This allows investigators to study or manipulate gene expression while avoiding double-strand breaks, supporting more controlled genome-engineering strategies.
Binding alone can regulate a locus in two ways described for this platform. The occupied dCas9 complex may physically block transcription, or it may serve as a position-specific platform for recruiting regulatory effector proteins. These alternatives let the same targeting logic support either direct interference with transcription or active control through an attached regulator.
Unlike DNA-cutting CRISPR systems, this complex is designed to alter gene activity without introducing double-strand breaks. Its effects arise from occupancy at a selected sequence or from regulatory proteins recruited there, rather than from DNA cleavage. This distinction makes it useful when controlled expression changes are preferred.
At a conceptual level, a CRISPR-dCas9 experiment begins by choosing a single-guide RNA complementary to the genomic region of interest. The guide directs dCas9 through RNA-DNA base pairing, after which the bound complex is used either to obstruct transcription or to recruit a regulatory effector. The selected mode determines whether the study emphasizes repression, activation, or regulatory control.
These complexes are useful when a study asks how changing the activity of a selected gene or genomic region affects a cell. Targeted repression or activation can be applied in functional genomics and cellular pathway analysis, while epigenetic regulation and locus visualization extend the platform to additional forms of biological investigation.