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NitAn Biotech LLC

NitAn Biotech LLC
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Viral Vector generation, cDNA libraries, cloning shRNA
Viral Vector generation, cDNA libraries, cloning shRNA
 

Viral Vector Services


Viral generation requires in depth knowledge in a wide variety of fields like molecular biology, cell biology, virology and biochemistry. Our scientists have successfully constructed efficient viruses in the past. The key features of our services include cDNA libraries (human, rat and mouse), fast turnaround time, diverse vector system and full consultation. Our construction procedure includes, cloning of your gene of interest or shRNA into the viral vector of customer’s choice, producing low or high titer viruses, amplifying the viral stock and purification/titer determination. Experience in viral vector services makes us unique in this area of service. According to the need of gene expression based on the limitations and advantages, we can provide different tools like adenovirus (AdV), lentivirus (LV) and adeno-associated virus (AAV) for our customers. Below we have summarized important features to consider;

Viruses

Packaging Capacity

Inflammatory Response

Advantage

Disadvantage

AdV 8-40 kb High 1. Infects all cell type (primary, dividing & non-dividing) with ~100% gene delivery efficiency
2. Does not integrate with host genome
3. Largest gene cloning capacity (up to 7.5kb)
High inflammatory response compare to AAV
LV 8 kb Low 1. Infects both dividing and non-dividing cells with 30% efficiency
2. Recombinant LV is easy to develop
Risk of host genome integration and potential activation of oncogenes
AAV ~ 4.7 kb Low 1. Infects all cell type
2. Low inflammatory response compare to AdV
3. Non-pathogenic
4. Gene cloning capacity 3 kb
Small packaging capacity

Viral Vector Services


You do

Our Scientists do

  1. Select the viral vector
  1. Helping in selecting appropriate tool
  2. Generate the gene of interest (GOI)
  3. Clone in GOI into viral vector
  4. Packaging virus
  5. Amplification & Purification
  6. Titer determination & Quality control

Adenoviral Vector Services

Adenoviral Vector ServicesThe adenovirus (AdV) is a non-enveloped, icosahedral particle containing linear double-stranded DNA. AdVs infect both non-dividing and dividing cells and replicate as episomal elements in the nucleus. There are 51 immunologically distinct adenoviruses exists. The majority of the adenovirus vectors are developed based on serotypes 2 and 5 by replacing either the E1 or E3 gene with a transgene, resulting in the loss of viral replicability. These recombinant viruses are replicable only in cells (like HEK) that express the E1 or E3 gene products at very high concentrations, creating a highly efficient, controllable viral vector system for experimental and clinical applications.

AdV construction, production, preparation and characterization

NitAn Biotech uses commercial adenovirus vectors (as per customer’s choice) for recombinant AdV construction, which are standard E1- and E3-deleted. We offer cloning in plasmids which expresses the transgene as the single gene product or simultaneously with fluorescent reporter gene, for analyzing the transfection efficiency. We also offer cloning in plasmids which have controllable gene expression like Tet system or Doxycyclin. Recombinant adenovirus vectors are verified by restriction digestion and sequencing, before transfection. Linearized adenoviral vector plasmids are transfected to HEK 293 cells to obtain AdVs. After multiple rounds of amplification, AdVs are purified and titered before mailing to customers. We also offer ready-to-use AdVs with no transgene or with GFP/LacZ reporter genes that can be used as controls.

Lentiviral Vector Services

Lentivirus (LV) offers great advantage over the adeno- or reteroviruses in delivering genes to quiescent and dividing cells. LVs integrate into host genome and being used for transient and stable gene expression. LVs have been successfully used to express platelet-derived growth factor (PDGF), platelet-factor VIII in diabetic and hemophilia mouse models, correcting the defective genes. NAB procures LV expression plasmids of customers choice (Gateway system from Invitrogen, Lenti-X HT Packaging System from Clontech) to generate LVs. Our LV service platform includes titering and sterility assay.

Adeno-Associated Viral Vector Services

Adeno-associated virus (AAV) is an excellent tool for gene therapy which infects humans and some other primate species. AAVs causes very mild immune response, thereby it overrides the use of other gene delivery tools like adenovirus or retroviruses. AAV infects a variety of cell types and expresses the gene of interest sustainably. AAV integrates into human genome at a specific site in the 19th chromosome. To date, AAVs are used in different phases of clinical trials for different diseases, particularly, AAV2 is reported to kill cervical, breast, prostate and squamous cell cancer cells, without affecting healthy cells. NAB offers AAV2 amplification, purification and titer determination.

For quote requests or orders ; please contact us or fax 1-614-573-7457 with a description of the project requirements. Orders with purchase order will be processed immediately. Please submit DNA or viral inoculums along with hard copy of the form with additional information.
 

OUR SERVICES

 
Viral Vector generation, cDNA libraries, cloning shRNA
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100 Science Village 1381 Kinnear Road Columbus, OH-43212 Tel: 614 893 3137 Fax: 614 573 7457