Shrna Expression
Shrna Expression - 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.
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).
09:51
Generation of Stable Human Cell Lines with Tetracycline-inducible (Tet-on) shRNA or cDNA Expression
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Cited by 16 •
2013
A rapid and simple way to generate human cell lines with inducible and reversible cDNA overexpression or shRNA-mediated knock-down of the gene of interest. This method enables researchers to reliably and highly reproducibly manipulate cell lines that are difficult to alter by transient transfection methods or conventional knockdown/knockout strategies.
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.
09:12
MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
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Cited by 11 •
2011
Here we use a human LentiPlex pooled library and traditional sequencing methods to identify gene targets promoting cell survival. We demonstrate how to set up and deconvolute a LentiPlex screen and validate the results.
FAQs
After transcription from the introduced DNA construct, the short hairpin RNA undergoes processing into small interfering RNA. This processed product is then loaded into the RNA-induced silencing complex, or RISC. These steps convert the original hairpin transcript into the form that can guide recognition of a complementary messenger RNA and initiate gene-silencing activity.
Sequence complementarity directs the silencing machinery toward the messenger RNA produced by the selected gene. Once the processed small interfering RNA guides RISC to a matching transcript, the messenger RNA can be degraded or its translation can be suppressed. Consequently, the target sequence determines which gene is affected and supports targeted analysis of gene activity.
The introduced DNA construct provides the template from which cells transcribe the hairpin RNA. Because the construct initiates production of the silencing precursor inside the cell, it supports targeted and sustained knockdown rather than relying only on a single externally supplied RNA molecule. This feature makes the approach useful for examining gene activity over an experimental period.
A typical experiment begins by selecting the gene whose activity will be reduced and introducing a DNA construct designed to produce the corresponding hairpin RNA. Cells then transcribe and process the hairpin, load the resulting small interfering RNA into RISC, and direct the complex toward complementary messenger RNA. The resulting reduction in gene activity supports functional investigation.
Researchers apply shRNA expression when they need to reduce the activity of a selected gene and examine the resulting biological consequences. In cultured cells and model organisms, targeted knockdown can help investigate gene function, explore cellular pathways, and connect a gene with a particular process. The approach is therefore useful for testing functional relationships within biological systems.
By reducing activity from a selected gene, shRNA expression can help researchers model aspects of disease mechanisms and examine how cellular pathways respond to altered gene function. The same targeted strategy can contribute to evaluating potential therapeutic targets by showing whether changing a gene affects a biological process of interest in cultured cells or model organisms.