Tet-On System

Harness the power of inducible gene expression with LipExoGen’s Tet-On System, offering tailored lentiviral vectors for precise control over your gene studies. Our services include custom production of lentiviral particles for shRNA, cDNA, and CRISPR-Cas9 sgRNA, utilizing our innovative Tet-On vectors for exceptional research outcomes.

Service Guide

Option 1: shRNA Expression System

  • Description: Custom lentiviral vectors for inducible shRNA expression. Ideal for gene silencing with the added control of doxycycline induction.
  • Applications: Gene knockdown studies, functional genomics, and therapeutic target validation.

Option 2: cDNA Expression System

  • Description: Tailored for overexpression of your gene of interest via inducible cDNA vectors. Leverage controlled gene activation to dissect function and mimic disease models.
  • Applications: Overexpression studies, pathway analysis, and disease modeling.

Option 3: CRISPR-Cas9 Knockout System

  • Description: Customizable sgRNA vectors for CRISPR-mediated gene editing with inducible expression for precise genomic alterations. Cas9
  • Applications: Gene editing, functional genomics, and development of gene therapies.

Advantages of Tet-On System

  • Inducible expression offers unparalleled control over gene activity, essential for dynamic studies and therapeutic development.
  • Simplifies experimental design by using a single cell population, enhancing reproducibility and accuracy.
  • Our vectors are designed for high-efficiency delivery and expression, suitable for a wide range of cell types.

Case Study

We have consistently offered our clients a specialized service for producing recombinant antibodies within their specified cells. The case study below exemplifies our process, focusing on a customer who was interested in rapidly obtaining purified recombinant anti-CD33 antibodies utilizing their proprietary DNA sequence.


Antibody-Producing Cell Screening:

The primary objective at this stage is to isolate high-capacity antibody-producing cells. After infection with high-titer lentivirus produced using the customer’s DNA sequence, the transduced cells underwent puromycin selection. Colonies of infected cells were then chosen specifically to identify those with high antibody-producing capacities. We use the culture medium from each cell colony and screen it with Western-blot (Figure 3A). Typically, after screening 25 to 30 colonies, we can establish a stable, high-producing cell line.

Purification of rAb and Gel Image Analysis:

Once we’ve identified high producing cells, the next step is the purification of the antibody. The success of this purification is then verified through a gel image stained with Coomassie blue, emphasizing the purified rAb after its treatment with DTT (Dithiothreitol) (Figure 3B). 

Functional Validation:

In this case, we produced a recombinant anti-human CD33 antibody. To assess its functionality, the antibody was fluorescently labeled and used to stain human AML cells provided by the client. The results confirm the functionality of the recombinant anti-CD33 antibody, given its robust binding affinity to the cells, which is known to exhibit surface CD33 expression based on separate staining with commercially available fluorescent anti-CD33 antibodies.

Figure 3. Snapshots of the validation procedure for recombinant antibody development using the Lenti-mAb system        A. Antibody-producing cell screening. Cell culture medium from individual colonies of the lentivirus-transduced and puromycin-resistant HEK cells are analyzed by Western blot to identify those with high expression of the recombinant antibodies. B. Reducing SDS-PAGE with Coomassie blue stain is performed to verify successful purification of the expressed recombinant antibody. C. The functionality of the purified recombinant antibody was assessed by fluorescently labeling with Alexa Fluor 488, followed by immunofluorescence staining the human CD33 expressing AML cells provided by the client. D. Further functional validation was demonstratred using the fluorescently-labeled antibody using flow cytometry with the clients’ human AML cells.

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