Shrna Cell Lines
Shrna Cell Lines - 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.
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.
01:16
Cell Lines
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2023
A cell line is a population of cells grown in vitro that can be subcultured over several generations. Normal cells cease to divide after a certain number of cell divisions, a process known as replicative senescence. This number, called the Hayflick limit, was conceptualized by Leonard Hayflick in 1961 when he observed that fetal cells grown in culture could only divide 40-60 times. This limit is due to the shortening of the telomeres during each round of cell division, preventing cell division...
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.
11:39
Chromatin Immunoprecipitation (ChIP) in Mouse T-cell Lines
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Cited by 13 •
2017
This work describes a protocol for chromatin immunoprecipitation (ChIP) using a mature mouse T-cell line. This protocol is suitable to investigate the distribution of specific histone marks at specific promoter sites or genome-wide.
FAQs
After shRNA is produced inside the cell, cellular processing converts it into small interfering RNA (siRNA). The siRNA associates with the RNA-induced silencing complex (RISC), which uses sequence complementarity to recognize the selected messenger RNA. RISC then promotes degradation of that transcript or blocks its translation, reducing the amount of protein produced from the targeted gene.
Sequence complementarity determines which messenger RNA is targeted: the processed siRNA guides RISC toward a matching transcript rather than acting broadly across the cell. Silencing can therefore arise through two related outcomes, transcript degradation or inhibition of translation. This distinction matters when interpreting experiments because reduced gene expression reflects altered messenger RNA or its translation, not removal of the gene itself.
Compared with transient RNA interference, stable shRNA cell lines maintain gene suppression across extended experiments. That persistence allows researchers to observe phenotypes that may take time to develop, rather than limiting analysis to a short intervention window. The stable format also supports reproducible knockdown between experiments, which is important when comparing cellular responses to the same genetic perturbation.
Creating a model begins with cultured cells engineered to produce shRNA directed against a selected gene. Once the cellular machinery processes that shRNA, the resulting silencing pathway lowers expression of the target. The model can then be maintained as a stable cell line and examined across experiments, providing a consistent setting for studying consequences of reduced gene expression.
Researchers apply shRNA cell lines to test gene function by observing how cells change when expression of a selected gene is reduced. The same models can be used to examine signaling pathways, disease mechanisms, and drug responses. Because suppression persists, they are especially suited to experiments in which the relevant phenotype or treatment response emerges over an extended period.
Interpretation centers on phenotypes or responses associated with reduced expression of the target gene. In biology, these outcomes can connect a gene to a signaling pathway, disease-related mechanism, or response to a drug. Comparing results across repeated experiments is aided by stable knockdown, which helps identify reproducible effects when investigating cellular consequences over time.