Expression Negative Control Lentivirus (EF1A Promoter/Hygromycin, Puromycin, or G418)

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82212

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Description

Expression Negative Control Lentivirus (EF1A Promoter/Hygromycin) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viral particles do not express a specific protein but include only a geneticin, hygromycin, or puromycin selection marker under the control of the EF1A promoter (Figure 1).

Figure 1. Schematic of the lenti-vector used to generate the Expression Negative Control Lentivirus (EF1A Promoter/G418). This is a SIN vector.

Figure 1. Schematic of the lenti-vector used to generate the Expression Negative Control Lentivirus (EF1A Promoter/Hygromycin). This is a SIN vector.

Figure 1. Schematic of the lenti-vector used to generate the Expression Negative Control Lentivirus (EF1A Promoter/Puromycin). This is a SIN vector.

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Product Data Gallery

Luciferase activity in HEK293 cells transduced with Expression Negative Control Lentivirus and Firefly Luciferase Lentivirus. (G418)

Luciferase activity in HEK293 cells transduced with Expression Negative Control Lentivirus and Firefly Luciferase Lentivirus (Hygromycin)

Luciferase activity in HEK293 cells transduced with Expression Negative Control Lentivirus and Firefly Luciferase Lentivirus. (Puromycin)

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer ≥10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
Thaw Medium 1	BPS Bioscience #60187
Firefly Luciferase Lentivirus (EF1A Promoter/Geneticin, Hygromycin, or Puromycin)	BPS Bioscience #78740
Lenti-Fuse™ Polybrene Viral Transduction Enhancer	BPS Bioscience #78939
96-well tissue culture-treated assay plates
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690

Formulation

The lentivirus particles were produced in HEK293T cells in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.

Background

The use of antibiotics in cell culture as a method to establish stable cell lines by imposing selective pressure on cells expressing the antibiotic resistance, is a common practice. However, the use of antibiotics can result in altered gene expression and regulation. For instance, analysis of the impact of the use of penicillin-streptomycin has identified 209 penicillin-streptomycin specific genes, including transcription factors. In view of these findings, care should be taken when performing genetic, genomic and biological studies in cells treated with antibiotics. In those cases, the use of a control where only the antibiotic resistance is expressed can prove valuable in assessing the specificity of the cellular response to the expression of a protein of interest

References

Ryu A., et al., 2017 Scientific Reports 7:7533

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Applications

Negative control in experiments involving transduction.
Generate cell pools following geneticin, hygromycin, or puromycin selection, to validate the specificity of the effects observed by expression of a protein of interest.

Shipping Temperature

-80°C

Notes

To generate an Expression Negative Control stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of G418 (as predetermined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, following by clonal selection.

Biosafety
The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only

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Expression Negative Control Lentivirus (EF1A Promoter/Hygromycin, Puromycin, or G418)

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