Lentivirus Packaging Services

Ensuring efficient, effective, safe, and reliable gene delivery

Overview

PackGene provides end-to-end lentiviral vector design and packaging services with superior quality driven by a blend of proprietary technologies and reagents. Our advances in lentivirus packaging optimize recombinant lentivirus production, especially for third-generation lentiviral vector systems. Benefits of this optimization include improvements in titer, purity, viability, and consistency in producing your custom lentivirus. Recombinant lentivirus is derived from modifications of the HIV-1 genome and stands as the most widely used viral vector for effective gene delivery in mammalian cells. In contrast to plasmid DNA vectors that only facilitate transient and episomal expression, lentiviral vectors enable permanent transgene expression by integrating foreign genes into the host cell’s genome. The design of a recombinant lentivirus involves both substituting a portion of the lentivirus genome with a target gene and enhancing the virus’s capability to infect a broad range of cell types, including neurons, liver cells, heart muscle cells, tumor cells, endothelial cells, and stem cells. The use of recombinant lentiviruses for transgene expression in mammals can be well tolerated and induces minimal immune responses. Thus, cell transfection with lentiviruses is appropriate for both in vitro and in vivo applications.

PackGene’s lentiviral products are versatile, with the capability to drive the expression of coding or non-coding genes like shRNA and microRNA. We provide 3rd-generation systems containing a modified 3’ LTR with a “self-inactivating nature”. This means that our 3rd-generation viruses are capable of infecting target cells but do not continue to infect other cells nor do they use host cells to produce new viral particles.

Highlights

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Efficient and prompt

delivery time as quick as 8 to 12 business days

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Accurate titer measurements

post-transduction qPCR eliminates any overestimation concerns

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Professional technical support

a response to any inquiries or requests will be delivered within 1 business day

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Enhanced Safety

“Self-inactivating” 3rd-generation system increases biosafety
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One-stop solution

from vector design and virus packaging to thorough analysis and testing

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Secure shipping

Dry ice and cold chain logistics to maintain optimal efficacy

No more concerns of overestimation

Accurate titer measurements by post-transduction qPCR

96-well plate
after transduction
96-well plate, 1.5E+04 cells/well, virus 1.8µL, 72hrs after transduction See more QC test data

Service Details

Service type Application Quantity  Concentration  Turnaround time
In vitro grade Cell Culture 5E7 TU ≥1E+8TU/mL 8-10 business days
1E8 TU
5E8 TU
1E9 TU
1E9~1E12 TU Inquiry
In vivo grade Cell Culture and in vivo 5E7 TU ≥1E+8TU/mL 15-20 business days
1E8 TU
5E8 TU
1E9 TU
1E9~1E12 TU Inquiry
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  • Yield is not guaranteed for constructs with elements detrimental to the viral packaging process. This includes but is not limited to, toxic genes, genes that disrupt viral packaging, genes that disrupt cell or lentiviral RNA integrity, sequences prone to rearrangement or secondary structures, nucleoproteins, transmembrane proteins, receptor genes, and gene segments with over 6.5kb between LTRs.

Technical Details

lentivirus workflow

Quality Control

Category QC test Description Release standard
Titer Post-transduction  qPCR Infection of cells followed by quantitative qPCR to determine the titer of lentiviruses
Report transduction unit
Transduction test Gradient dilution of infected cells and count of fluorescent cells
Provide bright field and fluorescent microscopy images
p24 Elisa* ELISA test for the p24 protein, a core capsid protein of HIV that is used to measure lentivirus titer
Report titer
Contamination Mycoplasma test* Measure mycoplasma DNA by PCR Guaranteed negative
Bioburden* Plate count method: Quantification of live microorganisms (fungi, bacteria, etc.) using colony-forming units (CFUs) No growth on the agarose plate
Safety Endotoxin LAL* Measure endotoxin level by gel clot assay Report

* QC with additional charges

Learn more about our lentivirus QC tests

Post-transduction qPCR: This test uses quantitative polymerase chain reaction (qPCR) to determine the amount of viral RNA present in cells after they have been transduced with lentivirus. This assay is used to determine the efficiency in which a lentivirus delivers its genetic material into host cells.

Transduction test: In this test, a sample of cells are infected with the lentivirus, and transfection efficacy is determined by measuring either transgene expression or the expression of a reporter gene such as GFP (Green Fluorescent Protein). This assay is used to evaluate the efficiency with which the lentivirus drives transgene expression in target cells.

Bright-field Fluorescent field   Bright-field Fluorescent field
96-well plate, 1.5E+04 cells/well, virus 18µL, 72hrs after transduction 96-well plate, 1.5E+04 cells/well, virus 1.8E-03µL, 72hrs after transduction
96-well plate, 1.5E+04 cells/well, virus 1.8µL, 72hrs after transduction 96-well plate, 1.5E+04 cells/well, virus 1.8E-04µL, 72hrs after transduction
96-well plate, 1.5E+04 cells/well, virus 1.8E-01µL, 72hrs after transduction 96-well plate, 1.5E+04 cells/well, virus 1.8E-05µL, 72hrs after transduction
96-well plate, 1.5E+04 cells/well, virus 1.8E-02µL, 72hrs after transduction 96-well plate, 1.5E+04 cells/well, virus 1.8E-06µL, 72hrs after transduction

p24 ELISA: The p24 antigen is a component of the HIV virus particle and can be used as a marker for virus quantity. In this test, ELISA is used to measure the concentration of p24 viral particles in a lentivirus preparation. This assay is used to quantify the titer of a lentiviral sample.

Mycoplasma test: Mycoplasma contamination is a common and serious issue in cell culture and can substantially reduce cell growth and lentivirus production. A mycoplasma test is conducted to ensure that the cell cultures used in lentivirus production are free from mycoplasma contamination.

Bioburden: This test measures the total number of living microorganisms present in the lentivirus product or on the surfaces of production equipment. The bioburden test is important for assessing the sterility and overall cleanliness of the production process.

Endotoxin LAL: The Limulus Amebocyte Lysate (LAL) test is used to detect and quantify bacterial endotoxins, which are components of the outer membrane of Gram-negative bacteria. Endotoxins can be potent immunostimulatory molecules, and their presence in lentivirus preparations can induce inflammatory responses in patients. Therefore, the Endotoxin LAL test is crucial for confirming that the lentivirus product is safe for in vivo applications or clinical use.

Lentiviral Transfer Plasmid

PackGene, a prominent provider in the field of gene therapy, offers a diverse range of lentiviral vectors that have been designed for several gene manipulation applications. Available vectors are tailored to facilitate gene overexpression or to drive the expression of shRNA.

Gene Overexpression Lentiviral Vectors: These vectors are engineered to increase the expression of a specific gene within a target cell. They are commonly used to study the function of a gene, to model a disease, or in various therapeutic applications.

PackGene’s overexpression vectors contain a strong promoter to drive high levels of transgene expression. We will first clone your gene of interest into any of our pre-designed vectors before packaging that vector into lentivirus. Following transduction, your gene will be integrated into the host cell’s genome, which results in sustained expression of the gene. This approach is valuable for understanding gene function, protein interactions, and potential therapeutic interventions where increased expression of a particular gene may be beneficial.

PackGene’s ready-to-use LVV transfer plasmid backbones are listed below. You may also design your own LVV transfer plasmid, and we will synthesize it for you.

Vector ID Promoter GOI capacity Promoter for Selection marker Selection marker
XL03C CMV 3.3Kbp EF1 CopGFP-2A-Puro
XL10C CMV 4Kbp EF1 mRuby2
XL11C CMV 4Kbp EF1 mCitrine
XL12C CMV 4Kbp EF1 EYFP
XL22C CMV 3.2kp EF1 mCherry-2A-Neo
XL23C EF1 4.2kb T2A CopGFP

shRNA Lentiviral Vectors: Short hairpin RNA (shRNA) vectors are used for gene silencing applications. PackGene’s shRNA vectors are designed to express shRNA sequences that are processed into small interfering RNA (siRNA) within the cell. These siRNA molecules guide cellular machinery to target mRNA to facilitate mRNA, degradation and thus reduce expression of the target gene. shRNA lentiviral vectors are crucial tools in gene function studies, allowing researchers to knock down the expression of specific genes and to observe the resulting phenotypic changes. They are also used in therapeutic research, particularly in diseases where the downregulation of a gene product may be beneficial.

Vector ID Promoter Promoter for Selection marker Selection marker
XL03B U6 promoter EF1a EGFP-2A-Neo
XL04B U6 promoter CMV CopGFP-2A-Puro
XL05B U6 promoter CMV Puro
XL06B U6 promoter PGK Puro-lRES-mCherry
XL07B H1 promoter CMV CopGFP
XL08B H1 promoter CMV CopGFP-2A-Puro
XL09B H1 promoter CMV Puro
XL012B U6 promoter CMV Neo

miRNA Lentiviral Vectors: MicroRNA (miRNA) are small, non-coding RNAs that play critical roles in regulating gene expression at the post-transcription level. PackGene’s miRNA lentiviral vectors are engineered to overexpress or inhibit the activity of specific miRNAs. They enable the study of miRNA function in various biological processes and diseases, offering insights into miRNA-mediated regulation of gene expression. These vectors are valuable for both basic research and in the development of miRNA-based therapeutic strategies. Manipulation of miRNA function may be especially revealing in areas like cancer, where miRNAs are known to play pivotal roles in disease progression and the response to treatment.

To design a miRNA LVV transfer plasmid, we recommend a H1 or U6 promoter followed by your miRNA sequence, then include a strong promoter and a selection marker as needed.

Resources
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Service Flyer

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Lentivirus Infection Instructions

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