• Virus Packaging (Lentivirus and Retrovirus)

Virus Packaging (Lentivirus and Retrovirus)

Applied StemCell provides lentiviral and retroviral custom virus packaging services for your CRISPR/Cas9 components, CAR-T expression vectors and other transgenes, for efficient delivery into cell lines (such as hard-to-transfect cell lines, primary cell lines) as well as direct injection into animal models.

  • Rapid, custom lentivirus or retrovirus virus packaging
  • Ready-to-transduce viral particles provided to you
  • High quality, high titer viral particles in 10 days
  • VSV-G pseudotyped viruses that exhibit broad tropism across a range of cell types
  • High titer amplification of viruses, up to 10^9 infectious units per ml
  • Accurate viral titers quantified by qRT-PCR
  • Confirmation by DNA sequencing
  • Large scale production, clonal expansion and cryopreservation
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Application Notes

Chimeric Antigen Receptor (CAR)-T Immunotherapy is a revolutionary T cell-based therapy to treat various types of cancer with a high success rate. CAR-T cell therapy is a type of adoptive cell transfer (ACT) which uses synthetic chimeric antigen receptor expressed on genetically engineered autologous T cell from PBMCs of cancer patients, to recognize and bind tumor-specific antigens on surface of cancer cells.1,2 With two, FDA-approved CAR-T cell therapies in the market, Kymriah™ and Yescarta™, there is a heated race to develop efficacious CAR-T cell with better safety profiles and other ACT-based therapies such as tumor-infiltrating lymphocytes (TIL), T cell receptor gene therapy (TCR), for not just cancer but other immune-based disorders such as autoimmune diseases. 2-5 However, these primary immune cells (primary T cell, B cells and NK cells) are notoriously difficult to engineer, and scientists often rely on pseudo-lentiviral or retroviral-based transduction of targeting vectors to genetically modify the cells. To that end, Applied StemCell, a leader genome editing technologies and animal/ cell line model engineering, offers its molecular cloning expertise as a custom service to package your ACT/ CAR-T expression vectors, including CRISPR-based targeting vectors into high quality, high titer lentiviruses to efficient transduction of for preclinical research applications. You can now enjoy the results of your research without the stress to get you there.

Example of Gene Insertion in Primary T Cells Using Lentivirus

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Figure. Human primary T cells were infected with lentivirus encoding a mouse gene. Both non-transduced and transduced T cells stained with antibodies against CD3 (a T cell marker) and mouse gene. FACS analysis demonstrates that transduction efficiency of T cells is up to 80%.

References:

  1. Sadelain, M. (2017). CD19 CAR T Cells. Cell, 171(7), 1471.
  2. National Cancer Institute. (2017). CAR T Cells: Engineering Patients’ Immune Cells to Treat Their Cancers. https://www.cancer.gov/about-cancer/treatment/research/car-t-cells
  3. Zheng, P. P., Kros, J. M., & Li, J. (2018). Approved CAR T cell therapies: ice bucket challenges on glaring safety risks and long-term impacts. Drug discovery today, 23(6), 1175-1182.
  4. The Scientist Staff. (2018). Cell and Gene Therapy Tracker: Global CAR T-Cell Trials. https://www.the-scientist.com/infographics/cell-and-gene-therapy-tracker-64450
  5. Kansal, R., Richardson, N., Neeli, I., Khawaja, S., Chamberlain, D., Ghani, M., ... & Marion, T. (2019). Sustained B cell depletion by CD19-targeted CAR T cells is a highly effective treatment for murine lupus. Science translational medicine, 11(482), eaav1648.
Technical Details

Lentivirus and retrovirus transfection are some of the most efficient methods of mammalian gene delivery for in vitro and in vivo applications. Choose Applied StemCell’s custom virus packaging service to reduce stress and effort to design and prepare your viruses, and to advance your research with reliable and reproducible results.

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Do you have a retro/lentivirus plasmid vector you want to have packaged and scaled up in a virus for in vivo or in vitro applications? Just ship your vectors and we will perform scale up for the size you want.

Applied StemCell also offers a full service CRISPR vector design, construction and validation.

Ask us for details.

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