Overexpression Vector Construction

Gene overexpression involves delivering the target gene into host cells, resulting in its elevated expression. Gene overexpression is widely applied in gene function studies, disease model development, drug screening, protein production, and gene therapy.

Service Details

Services Lentiviral Stable Overexpression Vector Construction / Transposon-Based Stable Overexpression Vector Construction / Transient Overexpression Vector Construction / Custom Vector Construction for Special Requirements
Deliverables 1. Plasmid map
2. Plasmid sequencing results
3. Plasmid amplification instructions
4. Plasmid
Turnaround/Price   Consult online for details
Based on years of experience in cell biology, EDITGENE has developed a mature system for constructing gene overexpression vectors. We offer both stable and transient overexpression vector construction services. Using lentiviral transduction, we can efficiently integrate exogenous genes of up to 5 kb into the host cell genome, ensuring stable expression even after multiple passages. Additionally, for genes larger than 5 kb, we utilize transposon-based methods to achieve long-term stable expression in cells

EDI-Service Advantages

Our lentiviral packaging system enables the stable integration of target genes into the genome of host cells, providing a more reliable tool for gene editing.                                  
Our optimized transposon system offers highly efficient delivery capabilities and a larger payload capacity, ideal for long-term stable expression of larger genes.

Service Types

Customized gene overexpression vectors can be designed and constructed based on client requirements and specific gene characteristics.
1 Lentiviral Stable Overexpression Vector Construction
2 Transposon-Based Stable Overexpression Vector Construction
3 Transient Overexpression Vector Construction
4 Custom Vector Construction for Special Requirements

Plasmid Map

 
Lenti Overexpression Lentiviral Plasmid Map
 
 
 
pPB Overexpression Transposon Plasmid Map

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: The lnc-CTSLP8 upregulates CTSL1 as a competitive endogenous RNA and promotes ovarian cancer metastasis

Ovarian cancer is highly lethal and poorly prognostic, primarily due to metastasis. Long non-coding RNAs (lncRNAs) play key roles in tumor progression, but their roles in ovarian cancer metastasis remain unclear. Researchers analyzed the expression of lnc-CTSLP8 in ovarian cancer using public databases (TCGA and GEO) and validated it through qRT-PCR. They constructed lnc-CTSLP8 overexpression and knockout cell lines using lentiviral vectors and the CRISPR/Cas9 system to analyze cell proliferation, migration, and invasion. In vivo studies used an ovarian orthotopic tumor mouse model to observe autophagy and EMT markers in cells, and RNA immunoprecipitation and dual-luciferase reporter gene experiments confirmed the interaction between lnc-CTSLP8 and miR-199a-5p. The results showed high expression of lnc-CTSLP8 in metastatic ovarian cancer, and its overexpression promoted ovarian cancer progression while enhancing autophagy and EMT. Mechanistically, lnc-CTSLP8 upregulates CTSL1 as a competitive endogenous RNA, exhibiting oncogenic effects. CTSL1 inhibitors and miR-199a-5p overexpression eliminated the effects of lnc-CTSLP8 overexpression. The study indicates that lnc-CTSLP8 acts as a ceRNA in ovarian cancer and is a potential therapeutic target.

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

When selecting a vector, consider the purpose of the experiment and the type of host cells. For example, plasmid vectors are commonly used for gene expression or amplification in bacteria, while viral vectors are more suitable for gene transfer in mammalian cells. Additionally, the vector's promoter, replicon, and antibiotic selection markers should be chosen based on specific requirements.
During vector amplification, Escherichia coli (E. coli) strains are typically used. The commonly used strain for most non-recombinant vectors is DH5α, which is suitable for most applications. For recombinant vectors, such as lentiviral vectors and transposon vectors, the Stbl3 strain can be used for amplification. Stbl3 is a specialized E. coli strain derived from HB101, which has a mutation in the recombinase gene recA13, effectively suppressing recombination of long fragment terminal repeat regions and reducing the likelihood of erroneous recombination.

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