Shrna Constructs
Shrna Constructs - 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.
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).
03:47
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.
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.
FAQs
After a shRNA construct is transcribed, its RNA folds into a stem-loop hairpin rather than acting as a simple linear transcript. Cellular processing converts this structure into a small interfering RNA-like molecule. That product guides the RNA-induced silencing complex, or RISC, toward messenger RNA with a complementary sequence, promoting cleavage or degradation and thereby reducing expression of the selected gene.
Sustained expression allows the silencing signal to remain available beyond a brief experimental window, making these constructs useful for examining long-term gene silencing strategies. In biomedical research, that persistence can support functional studies in which investigators follow the consequences of reducing a disease-associated gene or pathway over an extended period rather than assessing only an immediate response.
Complementarity directs the silencing machinery toward the intended messenger RNA, which supports targeted reduction of the corresponding gene. However, specificity cannot be assumed solely from the intended match because unintended off-target effects remain a central consideration. For medical research, this distinction is important when interpreting whether observed changes reflect the selected gene or unintended interactions within the cellular system.
They allow researchers to reduce expression of a selected gene and examine how that change affects disease-relevant biology. This makes them useful for functional studies, therapeutic-target validation, and pathway investigation. The approach has been applied in research on cancer, infections, and inherited disorders, helping investigators clarify how associated genes contribute to biological processes involved in these conditions.
Researchers can reduce expression of a disease-associated gene and then examine the resulting effects on the pathway or biological process under study. If silencing changes that process, the experiment provides functional evidence about the gene’s relevance as a potential therapeutic target. This application supports target evaluation before pursuing longer-term gene-silencing strategies or other biomedical investigations.
Cancer, infectious disease, and inherited-disorder research are specifically supported applications. In each area, reducing expression of a selected gene can help investigators study disease-associated mechanisms, test the importance of particular pathways, or evaluate whether a gene represents a promising therapeutic target. Sustained expression also makes the approach relevant to investigations of longer-term silencing strategies.
Delivery, specificity, and unintended off-target effects are important considerations. A construct must support gene silencing in the relevant cellular setting, while its effects should remain sufficiently focused on the intended messenger RNA. These issues influence how confidently researchers can attribute an observed biological change to the selected gene and must be considered when evaluating therapeutic targets or disease pathways.