Lentiviral Expression
Lentiviral Expression - Related Videos
02:50
Lentiviral Transduction of Neural Progenitor Cells for Epigenetic Regulation of Alpha-Synuclein Expression
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2025
Source: Tagliafierro, L., et al. Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models. J. Vis. Exp. (2019)This video demonstrates the lentiviral transduction of neural progenitor cells (NPCs) for targeted epigenetic modification of alpha-synuclein, a key protein in Parkinson’s disease. The protocol enables methyltransferase-mediated DNA methylation, reducing SNCA expression for studying gene regulation...
09:53
Specific Marking of HIV-1 Positive Cells using a Rev-dependent Lentiviral Vector Expressing the Green Fluorescent Protein
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2010
We have developed a lentiviral vector that possesses, in addition to the Tat-responsive LTR, the Rev-response element (RRE) that can regulate reporter gene expression in an HIV-1 Tat- and Rev-dependent fashion. The vector permits the specific detection of replicating HIV in living cells via the expression of GFP.
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.
08:46
Production of Lentiviral Vectors for Transducing Cells from the Central Nervous System
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Cited by 44 •
2012
In this protocol we describe production, purification and titration of lentiviral vectors. We provide an example of lentiviral vector-mediated gene delivery in primary cultured neurons and astrocytes. Our methods may also apply to other cell types in vitro and in vivo.
10:58
A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids
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Cited by 81 •
2015
In this video protocol we give a step by step explanation of lentiviral transduction in organoids of primary intestinal epithelium and of processing and downstream analysis of these cultures by quantitative RT-PCR, RNA-microarray and immunohistochemistry.
FAQs
The vector’s RNA genome is first reverse-transcribed into DNA after entry into the target cell. That DNA typically integrates into the host genome, creating a lasting template from which the selected gene can be expressed. This integration explains why lentiviral expression can support stable cell lines and longer-term studies of cellular behavior rather than only short-lived gene delivery.
A promoter is the regulatory element that drives production of the selected gene from the integrated vector DNA. Consequently, vector design must match the experimental question: researchers need to evaluate not only whether the gene is present, but also whether its expression level is appropriate for interpreting cellular behavior. This consideration helps reduce confounding from poorly controlled expression.
Engineered vectors remove viral genes required for replication while retaining the elements needed for gene delivery and expression. This separates the experimental goal of introducing a selected gene from viral functions that would otherwise support replication. The resulting design is important for controlled biological studies, because researchers can focus on how the introduced gene changes target cells rather than on replication-related activity.
At a high level, a lentiviral expression experiment requires an engineered vector carrying the selected gene and a promoter, followed by delivery into target cells. After entry, the vector RNA is converted into DNA and typically integrated. Researchers then evaluate expression and cellular behavior, using appropriate controls to distinguish gene-specific effects from effects related to the delivery system.
Controls help determine whether an observed change in cell behavior reflects the selected gene rather than the vector or delivery process. They also provide a reference for judging expression levels, which is essential when vector design or promoter activity could influence results. Including appropriate controls therefore strengthens interpretation of stable-cell-line and functional studies.
This approach is useful when researchers need sustained gene activity to examine biological processes over time. In biology, applications include generating stable cell lines, manipulating cells that are difficult to transfect, studying development, investigating disease mechanisms, and evaluating therapeutic strategies. The common outcome is a system in which changes in cellular behavior can be examined in relation to the introduced gene.