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Applications: • Negative control in experiments involving transduction.• Generate cell pools following hygromycin selection, to validate the specificity of the effects observed by expression of a protein of interest.
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
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 or hygromycin or puromycin selection marker under the control of the EF1A promoter.
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.
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.
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.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: BSL-2
References: Ryu A., et al., 2017 Scientific Reports 7:7533
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