Shrna Inhibition
Shrna Inhibition - Related Videos
03:33
Generating Conditional Knockdown Cells Using a Tetracycline-Responsive Lentiviral shRNA Expression System
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2026
Source: Kubala, M. H. & DeClerck, Y. A. Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment. J. Vis. Exp. (2017)This video demonstrates the generation of conditional knockdown cancer cell lines using a Tet-inducible lentiviral shRNA system. It outlines the steps for viral transduction, antibiotic selection, and doxycycline-induced gene silencing.
00:46
Feedback Inhibition
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2019
Biochemical reactions are occurring constantly in cells, converting starting substances to different products, usually with the help of enzymes that speed the reactions. Without enzymes, it would take far too long for most reactions to occur to be useful to the cell! Since enzymes help control the rate of reactions, their activity is regulated so that appropriate amounts of starting materials, intermediate metabolites, and products are maintained in the cell. Excessive build-up or depletion of...
03:47
Short Hairpin RNA-Mediated Gene Knockdown in iHSPCs In Vitro: A Lentivirus-Based shRNA Expression System Delivery into iHSPCs for Knockdown of Specific Gene Expression
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2025
In this video, we demonstrate a method to perform transduction of shRNA lentiviral vectors to obtain stable knockdown cell lines in immortalized hematopoietic stem and progenitor cells (iHSPCs).
11:15
Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome
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Cited by 2 •
2018
We report a small hairpin RNA (shRNA) and next generation sequencing-based protocol for identifying regulators of X-chromosome inactivation in a murine cell line with firefly luciferase and hygromycin resistance genes fused to the methyl CpG binding protein 2 (MeCP2) gene on the inactive X chromosome.
01:30
Enzyme Inhibition
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2025
Inhibitors are molecules that reduce enzyme activity by binding to the enzyme. In a normally functioning cell, enzymes are regulated by a variety of inhibitors. Drugs and other toxins can also inhibit enzymes. Some inhibitors bind to the enzyme’s active site, while others inhibit enzymatic activity by binding to other sites on the protein structure. Competitive inhibitors occupy the active site of enzymes, making them unable to accommodate the substrate. However, sufficiently high...
FAQs
The guide sequence determines which messenger RNA is recognized because it is complementary to the intended target. After the resulting small interfering RNA associates with the RNA-induced silencing complex, the complex is directed to that transcript. Recognition can then reduce target protein production by promoting messenger RNA degradation or by repressing translation, linking sequence choice to the observed loss-of-function phenotype.
The hairpin encoded by the DNA construct is an upstream form of the silencing signal rather than the final targeting unit. Cellular processing converts it into small interfering RNA fragments, which can load into the RNA-induced silencing complex. This processing creates the guide-containing machinery needed to find complementary messenger RNA, making it a critical step between construct delivery and suppression.
Persistent activity allows investigators to examine gene function over extended periods rather than only at an immediate time point. That feature is especially relevant when studying neural development, synaptic signaling, neurodegeneration, or behavior, where effects may emerge over time. It also supports loss-of-function experiments in neurons and glial cells that require longer observation.
An experiment begins by providing cells with a DNA construct encoding the selected short hairpin RNA. The construct supplies the sequence that is processed into small interfering RNA, after which the fragments enter the RNA-induced silencing complex. The complex then recognizes complementary messenger RNA and reduces production of the corresponding protein, creating a targeted loss-of-function condition.
In neuroscience, the method can be applied to neurons and glial cells to test how particular genes contribute to synaptic signaling, neural development, neurodegeneration, and behavior. Reducing one target's protein production creates a loss-of-function setting, allowing the resulting cellular or behavioral changes to be related to that gene's role within the neural system.
By selectively lowering production of a target protein, shRNA inhibition can reveal whether that gene influences a neural process or disease-relevant phenotype. Findings from loss-of-function studies may identify genes whose activity merits further investigation as potential therapeutic targets. The sustained nature of the approach is useful when the relevant neural changes develop during longer-term experiments.