Shrna Transduction
Shrna Transduction - Related Videos
14:51
Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype
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Cited by 2 •
2022
High-throughput RNA interference (RNAi) screening using a pool of lentiviral shRNAs can be a tool to detect therapeutically relevant synthetic lethal targets in malignancies. We provide a pooled shRNA screening approach to investigate the epigenetic effectors in acute myeloid leukemia (AML).
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).
09:12
MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
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Cited by 10 •
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.
01:16
Transduction
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2025
Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome are...
FAQs
Once the vector delivers DNA to a target cell, the shRNA cassette is transcribed into a hairpin RNA. Processing converts this precursor into small interfering RNA, after which it is loaded into RISC. The guide strand then recognizes complementary messenger RNA, enabling its sequence-specific degradation or suppression. This pathway links vector delivery to reduced gene expression.
Sequence complementarity gives the guide strand a defined target: it directs RISC toward messenger RNA containing the matching sequence. This makes the assay suitable for testing the effect of reducing a selected gene rather than merely introducing a delivery vehicle. Measuring resulting phenotypic effects helps connect altered expression with gene function.
An important experimental choice is whether the intended knockdown should be stable or transient. Stable and transient formats allow investigators to design experiments around the persistence of reduced gene expression. This choice should match the biological question and desired duration of silencing, including studies comparing phenotypes across cultured cells or experimental organisms under different designs.
Vector selection and controls are central to interpreting shRNA experiments. Careful vector selection supports the intended delivery approach, while controls provide a reference for changes not attributable to the intended silencing. Together, these elements help distinguish a phenotype associated with reduced expression from effects related to the delivery process itself in the assay.
A basic workflow begins with DNA encoding the selected shRNA, followed by its carriage in a viral vector. The vector is introduced into target cells, where the cassette is transcribed and processed. Researchers can then examine reduced expression and associated phenotypic effects, using controls to interpret whether delivery contributed to the result.
Researchers use this approach to test gene function by reducing expression and examining the resulting phenotype. In cultured cells, it supports studies of gene function; in experimental organisms, it extends analysis to organism-level effects. The strategy also helps model disease-related pathways, provided vector-related effects are considered when interpreting the observed outcome.