Aldevron Breakthrough Blog
Bringing CRISPR Into the CAR-T Process
April 24, 2024 by Patrick Paez, Ph.D.
Webinar Recap: Going Non-Viral: How Cell Therapy Manufacturing is Moving Away From Viral Vectors
Chimeric antigen receptor T, or CAR-T, cell therapy comes from a long history of progress in the field of immunology, starting with observations in the 1950s of hematopoietic stem cell transfers and graft versus tumor activity, leading to early-generation CAR-T cell therapies in the 1990s.
By and large, CAR-T cell therapy has shown clinical success with liquid tumors (blood cancers) but has experienced challenges with solid tumor disease types like carcinomas and sarcomas. There have been some advances in improving the biology of CAR-T cells to infiltrate solid tumors and maintain tumor-killing capacities, improving its potential to expand its cancer indications for the future.
However, there are concerns regarding the cost of manufacturing therapies. Timelines can be long and complex given current labor and technical requirements for producing viral vectors used in editing patient cells. In our most recent webinar, Going Non-Viral: How Cell Therapy Manufacturing is Moving Away From Viral Vectors, we discussed whether the possible solution for bringing down costs and reducing the timeline for delivery would be a manufacturing shift to non-viral vector methods.
In the webinar, John Zuris, Ph.D., Director of Editing Technologies at Editas Medicine, and Hyatt Balke-Want, MD, a postdoctoral researcher at Cologne University Medical Center, discussed new novel manufacturing methods for CAR-T cell therapies to improve accessibility, offering a history of success and potential future clinical applications for their approach.
During this webinar, Drs. Zuris and Balke-Want described how their CRISPR-based knock-in methods could help achieve 70-90% knock-in rates in clinically relevant cell types, including CAR-T cells. Their novel CRISPR knock-in approaches used selection systems to create purified and enriched edited cell populations.
Dr. Zuris discussed the CRISPR homology-directed repair knock-in method developed at Editas Medicine called SeLection by Essential-gene Exon Knock-in (SLEEK). In this method, essential genes like GAPDH are targeted by a nuclease, and the essential gene and target gene are knocked back in conferring survival, of high numbers of edited cells.
Dr. Balke-Want’s CRISPR knock-in method, called homology-independent targeted insertion (HITI) CRISPR EnrichMENT (CEMENT), generates a high number of knock-in cells via an alternative enrichment process using methotrexate. This chemotherapeutic agent kills unedited cells in culture unless they have successfully incorporated the dihydrofolate reductase gene included in the donor constructs. Use the link below to watch the webinar and learn more about the methods mentioned, and to request more information from our experts.
- Watch the webinar on demand
- Learn more about Aldevron’s CRISPR services
- Have questions or want to learn more about these methods? Contact Patrick Paez
- Have an idea for a topic? Let us know!
Patrick Paez serves as a technical marketing manager for Aldevron’s marketing team. He earned a bachelor's degree in Pre-Medicine and Psychology, and a Ph.D. in Immunology at Virginia Commonwealth University (VCU) in Richmond, VA. His Ph.D. training at VCU's Medical College of Virginia campus was in a translational science lab at the Massey Cancer Center, where he focused on developing immunotherapeutic interventions in the field of immuno-oncology for phase 1 clinical trials.