mCherry Lentivirus (Hygromycin or Puromycin)

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78932

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Description

mCherry Lentivirus (Hygromycin or Puromycin) 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 viruses contain mCherry driven by an EF1a promoter and a hygromycin or puromycin selection marker (Figure 1).

Figure 1. Schematic of the lenti-vector used to generate the mCherry Lentivirus (Hygromycin). This is a SIN vector.

Figure 1. Schematic of the lenti-vector used to generate the mCherry Lentivirus (Puromycin). This is a SIN vector.

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Product Data Gallery

Expression of mCherry in HEK293 cells transduced with mCherry Lentivirus (Hygromycin)

Expression of mCherry in HEK293 cells transduced with mCherry 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
Lenti-Fuse™ Polybrene Viral Transduction Enhancer	BPS Bioscience #78939
96-well clear-bottom tissue culture-treated assay plates
Flow cytometer or fluorescence microscope

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

mCherry is a monomeric red fluorescent protein derived from DsRed found in the sea anemones Discosoma. It belongs to the mFruit family of monomeric red fluorescent proteins, which are improved versions of mRFP1 (monomeric red fluorescent protein 1) in terms of brightness and photostability. The use of fluorescent proteins allows for direct visualization of transfected or transduced cells under a fluorescent microscope or analysis by flow cytometry. The use of lentiviruses to introduce mCherry is a convenient strategy that allows expression of the markers in almost all mammalian cells and to easily determine transduction efficiency.

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Growth Media

For best results, the use of validated and optimized media from BPS Bioscience is highly recommended. Other preparations or formulations of media may result in suboptimal performance.

Media Required for the Proposed Assay
Thaw Medium 1 (BPS Bioscience #60187): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin

Applications

Positive control in experiments involving transduction.
Optimize transduction assays and track reporter protein expression over time.
Generate cell pools or stable cell lines expressing mCherry following hygromycin or puromycin selection.

Shipping Temperature

-80°C

Notes

To generate an mCherry stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of hygromycin (as pre-determined 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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mCherry Lentivirus (Hygromycin or Puromycin)

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