Lentiviral particles for fluorescent whole-cell labeling

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

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Tracking of mammalian cells transduced with lentivirus expressing AmCyan1 fluorescent protein (Cat. # 0011VCT rLV.EF1.AmCyan1-9)

Tracking of mammalian cells transduced with lentivirus expressing AmCyan1 fluorescent protein (Cat. # 0011VCT rLV.EF1.AmCyan1-9).

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Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins

Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins.

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

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Various cell types can be transduced with Vectalys lentiviral particles, including primary and stem cell lines

Various cell types can be transduced with Vectalys lentiviral particles, including primary and stem cell lines. The suggested seeding concentration and MOI vary depending on the specific cell type to be transduced. When the recommendations in this table are followed, transduction efficiency is high for immortalized, primary, and stem cells, regardless of whether they are adherent or suspension cells.

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Overview of the process for generating high-titer lentiviral particles

Overview of the process for generating high-titer lentiviral particles. Through this proprietary, large-scale process Vectalys is able to produce very high-titer, high-purity lentiviruses ready for transduction of many different cell types. The high titer allows for manipulation of even hard-to-transduce cells, like primary cells or stem cells, and the exceptional purity prevents cytotoxicity.

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Comparison of the purity of lentiviruses generated with Vectalys’ method or a classic purification method

Comparison of the purity of lentiviruses generated with Vectalys’ method or a classic purification method. Lentiviral proteins were separated by SDS-PAGE and stained with Coomassie Blue. The high purity of Vectalys lentiviral particles is clear when comparing the two methods; even when Vectalys particles are loaded at a 10-fold higher amount, very little contaminating protein is seen. The ratio of total particles (physical particles) to functional particles (transducing units) is lower for Vectalys purified lentivirus compared to classic methods of purification, indicating higher purity. Data provided by Vectalys.

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Not all titers are the same! Comparison of cells transduced with 100 M

Not all titers are the same! Comparison of cells transduced with 100 M.O.I. of lentivirus provided by Clontech/Vectalys versus 100 M.O.I. of lentivirus provided by Vendor S. Each premade virus preparation expresses a green fluorescent protein when transduced. Clontech/Vectalys provides a functional titer, so every cell treated with our virus preparation was transduced with multiple copies of virus, leading to strong expression of the fluorescent protein. By contrast, the titer provided by Vendor S includes nonfunctional lentivirus particles as a large percentage of the titer so 100 M.O.I of their virus preparation is not sufficient to effectively transduce the cells.

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What is a functional titer? Most vendors of ready-made viral particles determine their titers using a simple particle count

What is a functional titer? Most vendors of ready-made viral particles determine their titers using a simple particle count. This count will include free capsid protein, nonfunctional and immature virions, as well as functional virus.The functional titer provided by Clontech and Vectalys is not just the total particle number, it is a measurement of the total number of virions capable of integrating into cells, measured by transducing HCT116 cells. In real terms, this means our viral preps contain up to 100-fold more infectious particles than preps from other vendors.

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Achieve 100% transduction efficiency even in cell types that are typically hard to infect

Achieve 100% transduction efficiency even in cell types that are typically hard to infect. Super-concentrated highly purified lentivirus prepared by Vectalys permits transduction at high M.O.I.

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

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Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins

Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins.

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

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Whole-cell labeling using Clontech's ZsGreen1 fluorescent protein delivered using premade lentivirus (Cat. # 0010VCT rLV.EF1.ZsGreen1-9); nucleus is stained using DAPI. ZsGreen1 is 2.5X brighter than EGFP

Whole-cell labeling using Clontech's ZsGreen1 fluorescent protein delivered using premade lentivirus (Cat. # 0010VCT rLV.EF1.ZsGreen1-9); nucleus is stained using DAPI. ZsGreen1 is 2.5X brighter than EGFP.

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Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins

Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins.

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

The Vectalys Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles expressing fluorescent protein genes for transducing mammalian cells. The expression of fluorescent protein genes is driven by an EF1α promoter. These VSV-G pseudotyped viruses are capable of infecting both dividing and non-dividing cells. The high purification level of these lentiviral particles allows transduction of primary cells and immortalized cell lines without inducing any toxicity to the cells, even at high multiplicity of infection (M.O.I.).

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Mammalian cells transduced with lentivirus (Cat. # 0012VCT rLV.EF1.ZsYellow1-9) expressing ZsYellow1 fluorescent protein

Mammalian cells transduced with lentivirus (Cat. # 0012VCT rLV.EF1.ZsYellow1-9) expressing ZsYellow1 fluorescent protein.

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Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins

Map schematics of expression cassettes contained in particles for whole-cell labeling with fluorescent proteins.