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Lentiviral Packaging

Lentivirus is a unique retrovirus capable of effectively integrating foreign genes into the host chromosome, enabling long-term stable expression. The advantages of lentiviral vectors include broad cell infectivity, sustained stable expression, low immunogenicity, high cargo capacity, and relative safety. These characteristics make lentiviral vectors highly valuable and promising for biomedical research and clinical applications.

Service Details

Services Knockout Lentivirus / Overexpression Lentivirus / Interference Lentivirus / Library Virus
Deliverables 1. Titer Report
2. Map
3. Lentivirus
Turnaround/Price   Consult online for details
EDITGENE offers a well-established lentiviral packaging platform capable of producing lentiviruses tailored to meet various experimental needs. Our lentiviral vectors are carefully optimized, utilizing a mature packaging system designed to enhance biosafety, viral transduction efficiency, and gene expression levels. We use cell-based titration methods to accurately determine the active viral titer, ensuring optimal conditions for downstream experiments.

EDI-Service Advantages

Efficient Packaging System
Optimized lentiviral packaging technology improves packaging efficiency and titer, providing more effective tools for scientific research experiments.
Scientific Titer Measurement Method
Titer is measured using the cell titration method, ensuring accurate measurement of active virus titer and improving experimental precision.
Extremely High Safety
The lentiviral vector system used is carefully optimized, and the virus packaging system is mature, ensuring excellent safety.
Experienced Team
The technical staff are highly professional and experienced, capable of packaging lentivirus suitable for various experiments based on customer needs. 

Service Types

We offer various lentiviral packaging services tailored to customer needs:
Knockout Lentivirus Delivers and integrates the CRISPR/Cas9 knockout system into the genome, enabling efficient knockout of target genes
Overexpression Lentivirus Delivers and integrates exogenous gene fragments into the genome for efficient and stable expression of gene fragments
Interference Lentivirus Delivers and integrates the shRNA interference system into the genome for long-term and stable knockdown of target genes
Library Virus Delivers CRISPR/Cas library plasmids to cells and integrates them into the genome, effectively creating gene-edited mixed cell pools

Delivery Standards

Gene Knockout Virus, Overexpression Virus, Interference Virus CRISPR/Cas9 Library Lentivirus

Titer Report

 Titer Report
Plasmid Map Plasmid Map

Operation Manual

 Library Lentivirus（ ≥1×106 TU/mL, delivery sizes：1×108 TU、5×108 TU、1×109 TU）
Lentivirus (Titer ≥ 1×106 TU/mL，1 mL)  

Workflow

 

Literature Review

Genome-Wide CRISPR-Cas9 Knockout and Transcriptional Activation Screening
CRISPR-Cas9 is a powerful genetic screening tool that can be used to gain deep insights into gene functions and regulatory networks, with significant potential for applications in disease-related gene research. To demonstrate how to conduct genome-wide CRISPR-Cas9 knockout and transcriptional activation screenings, Joung et al. cloned a custom sgRNA library into lentiviral vectors, packaged the virus particles in HEK293FT cells, and collected them for transducing target cells. This approach enabled large-scale CRISPR-Cas9 knockout and activation screenings, successfully identifying key genes associated with specific phenotypes.
 
Lentiviral Packaging Solutions for Screening Candidate Genes
Joung, Julia et al. “Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening.” Nature protocols vol. 12,4 (2017): 828-863.

Advantage and Characteristic

Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic

Reference Materials

Article Title: Exploring the role of CBLB in acute myocardial infarction: transcriptomic, microbiomic, and metabolomic analyses

Specific changes in gut microbiota and metabolites are associated with acute myocardial infarction (AMI), and CBLB may play an important role in this process. However, the specific mechanisms of these interactions have not been thoroughly studied, leading to significant gaps in understanding. This study aims to investigate the effects of CBLB intervention on AMI mice through transcriptomic sequencing, 16S rDNA, and untargeted metabolomic analyses. The research team employed a multi-omics integrated strategy, including the transfection of CBLB lentiviral packaging vectors into 293T cells, followed by intervention in AMI mice, and conducting pathological staining, fecal 16S rDNA sequencing, and serum untargeted metabolomics analysis. The results showed that CBLB intervention significantly reduced inflammatory cell infiltration and collagen fiber formation in the infarct area of the mouse heart and led to key changes in microbiota, metabolites, and differentially expressed genes (DEGs). This suggests that CBLB may play a significant role in the regulation of AMI. The study confirms the potential of differentially expressed genes, metabolites, and microbiota in AMI regulation after CBLB intervention, providing a basis for future exploration of the therapeutic applications of CBLB.

Article Title: SENP1-Mediated deSUMOylation Regulates the Tumor Remodeling of Glioma Stem Cells Under Hypoxic Stress

This study explores the impact of SENP1-mediated deSUMOylation on the malignant behavior of glioma stem cells (GSCs) under hypoxic conditions and its clinical significance. Under hypoxic conditions, HIF1α upregulation activates the Wnt/β-catenin signaling pathway in GSCs, supporting glioblastoma (GBM) growth. SENP1-mediated deSUMOylation stabilizes the expression of HIF1α and β-catenin, promoting tumor formation driven by GSCs. Downregulating SENP1 expression in GSCs using lentivirus-packaged SENP1 shRNA resulted in decreased proliferation and differentiation of GSCs, with a reduction in tumorigenesis.HIF1α-induced activation of Wnt/β-catenin is dependent on SENP1-mediated deSUMOylation, promoting GSC-driven GBM growth. The study suggests that targeting SENP1 may effectively enhance the therapeutic efficacy for GBM.

Selected Customer Resources

IF=50.5
Nature

Abstract:

To date, more than half of global hepatocellular carcinoma (HCC) cases occur in China, yet comprehensive whole-genome analyses focusing on HBV-related HCC within the Chinese population remain scarce. To address this challenge, researchers initiated the China Liver Cancer Atlas (CLCA) project, aiming to conduct large-scale whole-genome sequencing to unravel the unique pathogenic mechanisms and evolutionary trajectories of HCC in China.

The researchers performed deep whole-genome sequencing on 494 HCC tumor samples, with an average depth of 120×, alongside matched blood controls, providing a detailed genomic landscape of HBV-associated HCC. Beyond confirming well-known coding driver genes such as TP53 and CTNNB1, the study identified six novel coding drivers—including FGA—and 31 non-coding driver genes.

Additionally, the research uncovered five new mutational signatures, including SBS_H8, and characterized the presence of extrachromosomal circular DNA (ecDNA) formed via HBV integration, which contributes to oncogene amplification and overexpression. Functional validation experiments demonstrated that mutations in genes such as FGA, PPP1R12B, and KCNJ12 significantly enhance HCC cell proliferation, migration, and invasion.

These findings not only deepen our insights into the genomics of HCC, but also open up new potential targets for diagnosis and therapy. View details>>

Candidate driver landscape

 

IF=27.4
Advanced Materials

Abstract:

During the acute inflammatory phase of tendon injury, excessive activation of macrophages leads to the overexpression of SPP1, which encodes osteopontin (OPN), thereby impairing tissue regeneration. The CRISPR-Cas13 system holds great promise for tissue repair due to its unique RNA editing and rapid degradation capabilities; however, its application has been limited by the lack of efficient delivery methods.

To address this, the researchers systematically screened various cationic polymers targeting macrophages and developed a nanocluster carrier capable of efficiently delivering Cas13 ribonucleoprotein complexes (Cas13 RNPs) into macrophages. Utilizing a reactive oxygen species (ROS)-responsive release mechanism, this system specifically suppresses the overexpression of SPP1 in macrophages within the acute inflammatory microenvironment of tendon injury.

Experimental results demonstrated that this targeted delivery strategy significantly reduced the population of SPP1-overexpressing macrophages induced by injury, inhibited fibroblast activation, and alleviated peritendinous adhesion formation. Furthermore, the study elucidated that SPP1 promotes fibroblast activation and migration through the CD44/AKT signaling pathway, and that inhibiting this pathway effectively mitigates adhesion formation following tendon injury. View details>>

Schematic diagram illustrating immune microenvironment-activated mRNA editing strategies of macrophages for PA therapy

IF=12.8
Biomaterials

Abstract：

Spinal cord injury (SCI) is a severe disabling condition that causes permanent loss of sensory, autonomic, and motor functions. While stem cell therapies, particularly mesenchymal stem cells (MSCs), show great promise for SCI treatment, their limited regenerative capacity restricts their application in tissue repair. The researchers observed that extracellular vesicles derived from antler bud progenitor cells (EVsABPC) may carry bioactive signals that promote tissue regeneration. Accordingly, they isolated and engineered EVs from ABPCs for SCI therapeutic investigation.

The study found that EVsABPC significantly enhanced neural stem cell (NSC) proliferation, promoted axonal growth, reduced neuronal apoptosis, and modulated inflammation by shifting macrophage polarization from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype. Moreover, engineered EVsABPC modified with cell-penetrating peptides demonstrated improved targeting to the SCI lesion site, markedly enhancing neural regeneration and functional motor recovery. These findings highlight EVsABPC as a promising candidate for SCI therapy. View details>>

Graphical abstract

IF=11.3
Journal of Hazardous Materials

Abstract:

S-metolachlor (S-MET) is one of the most widely produced and applied herbicides in China. Owing to its chemical properties, it tends to persist in soil and easily contaminates surface and groundwater through leaching and runoff. This environmental persistence poses a serious threat to plant development and, through the food chain, to human health.

To address the limitations of current detection technologies and meet the growing demand for high-efficiency analytical tools, the researchers employed a mammalian expression system to generate recombinant antibodies targeting S-MET.

Building on the successful expression of these antibodies, they established a sensitive immunoassay for monitoring S-MET residues in various environmental water samples. The icELISA results showed that the recombinant antibodies retained the sensitivity, specificity, and biological activity of the original monoclonal antibodies, delivering accurate and reproducible detection in river water, agricultural runoff, and tap water. View details>>

Graphical abstract

 

IF=10.7
Biosensors and Bioelectronics

Abstract:

MicroRNAs (miRNAs) are a class of small non-coding RNA molecules that regulate gene expression by interacting with the mRNAs of target genes. Given their crucial role in the development and progression of various diseases, miRNAs have emerged as promising biomarkers for clinical diagnostics.

In this study, researchers established a novel detection platform, termed DBmRCA, which combines dumbbell probe-initiated multi-rolling circle amplification with the high-sensitivity signal output of CRISPR/Cas12a. This enzyme-free, isothermal method enables accurate quantification of miRNA within just 30 minutes.

Clinical validation revealed that the expression levels of miR-200a and miR-126 were significantly downregulated in lung cancer tissues, and results from DBmRCA were consistent with those obtained by conventional techniques. With its high sensitivity, rapid turnaround, and simplified workflow, the DBmRCA platform presents a reliable tool for miRNA detection and holds strong promise for early diagnosis and therapeutic monitoring of lung cancer. View details>>

Graphical abstract

FAQ

Lentivirus is a gene delivery tool that introduces exogenous genes into cells. By using tool cells such as 293T, lentiviral vectors carrying target DNA fragments are packaged into lentiviral particles with cell-infectious activity. This packaging process includes constructing lentiviral vectors, preparing packaging plasmids, culturing tool cells, transfecting plasmids, collecting viral particles, purifying and concentrating viral particles, and titration.
Lentiviruses have high transduction efficiency and the ability to maintain stable gene expression over the long term, making them particularly suitable for cell types that are difficult to transfect. Furthermore, lentiviruses can integrate exogenous genes into the host genome, ensuring prolonged gene expression.
4.Poor cell condition after lentiviral infection may be caused by various factors. Here are some possible reasons and corresponding solutions: 1.High viral titer: High titers of lentivirus may cause cytotoxicity, preventing normal cell growth. Solution: Lower the viral titer and conduct a series of dilution experiments to find a titer that effectively transduces without adversely affecting cell growth. 2.Poor cell condition: The health status of cells before infection can affect growth after infection. Solution: Ensure cells are in optimal condition for infection, for example, by changing to fresh culture medium 24 hours before infection and ensuring appropriate cell density. 3.Toxicity of gene expression mediated by the virus: The gene carried by the lentiviral vector may be toxic to the cells, affecting their growth. Solution: If possible, use a control vector to determine if the problem is related to gene expression, and select appropriate vectors or genes for research. 4.Excessive antibiotic selection pressure: If antibiotics are used to select transfected cells, excessive concentrations of antibiotics may inhibit cell growth. Solution: Optimize the antibiotic concentration and use gradient experiments to determine the optimal concentration.

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