Lentiviral Vector Purification
Lentiviral Vector Purification - Related Videos
03:07
Lentiviral Vector-Based Transduction of Rat Fertilized Embryos
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2026
Source: Koza, P., et al. Generation of Transgenic Rats using a Lentiviral Vector Approach. J. Vis. Exp. (2020)This video demonstrates microinjection-assisted delivery of lentiviral vectors into fertilized rat embryos to achieve stable transgene integration for the generation of transgenic rats.
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.
10:42
A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells
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Cited by 21 •
2017
We describe the production strategy of integrase-deficient lentiviral vectors (IDLVs) as vehicles for delivering CRISPR/Cas9 to cells. With an ability to mediate quick and robust gene editing in cells, IDLVs present a safer and equally effective vector platform for gene delivery compared to integrase-competent vectors.
02:30
Gene Delivery to Mouse Intestinal Organoids Using Lentiviral Vectors
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2026
Source: Van Lidth de Jeude, J. F., et al. Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids. J. Vis. Exp. (2015)This video describes the lentiviral transduction of mouse intestinal organoids from a crypt-like structure containing suspension. Using a basement membrane matrix as a scaffold and growth factor–supplemented media, lentiviral vectors are introduced to enable stable gene...
16:04
A Tetracycline-regulated Cell Line Produces High-titer Lentiviral Vectors that Specifically Target Dendritic Cells
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Cited by 6 •
2013
Here, we use retroviral transduction and concatemeric transfection to create a cell line that can express the components of a lentiviral vector (LV) in the absence of tetracycline. This LV encodes GFP and is pseudotyped with a glycoprotein, SVGmu, which is specific for a receptor on dendritic cells.
FAQs
The workflow exploits measurable differences between lentiviral particles and unwanted material. Clarification first removes cells and debris, while filtration separates according to size. Later concentration or chromatography can use density, charge, or binding behavior to enrich particles further. Because these operations select by different physical or chemical properties, combining them can reduce host-cell proteins, nucleic acids, and process reagents more effectively than relying on one step.
Nuclease treatment targets contaminating nucleic acids that remain after the culture material has been clarified. This step complements, rather than replaces, filtration or chromatography: nuclease reduces a particular contaminant class, whereas the other operations separate or enrich material using size, density, charge, or binding behavior. Including complementary mechanisms helps produce a preparation with improved purity for downstream transduction and characterization.
Concentration and chromatography serve different purification needs. Concentration enriches the vector from a processed volume, whereas chromatography provides a separation based on properties such as charge or binding behavior. The choice therefore depends on the desired balance among recovery, infectivity, purity, and workflow suitability. These quality attributes should be assessed together because increasing enrichment does not by itself guarantee the best functional preparation.
A typical workflow begins with clarification of the cell-culture material to remove cells and debris. Nuclease treatment may then be combined with filtration and a concentration or chromatography step, with the sequence selected to reduce contaminants while retaining functional particles. The resulting preparation is evaluated for recovery, infectivity, and purity. This staged approach links each operation to a defined separation or quality-control objective.
Process conditions are optimized by balancing competing outcomes rather than maximizing a single measurement. Recovery indicates how much vector is retained, infectivity reflects functional performance, and purity reflects removal of unwanted culture and process components. A workflow that improves one attribute while substantially reducing another may be unsuitable for the intended study, so these measures guide method selection and optimization.
In biology research, the purified preparation supports more consistent transduction experiments and functional characterization than a less controlled input. It can also contribute to preclinical development of gene-transfer strategies, where reproducible vector quality and biosafety are important. Purification is therefore not only a cleanup step; it connects cell-culture processing with interpretable biological results and later evaluation of lentiviral delivery approaches.