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CRISPR-edited CAR Cell Technology

Chimeric antigen receptor (CAR) T cells have shown great promise in the treatment of hematology and solid malignancies. However, despite the success in this field, there are still some major challenges, including accelerated T cell failure, potential toxicity, and insertional tumorigenesis. To overcome these limitations, the latest advances in clustering, regularly spaced, short palindromic repeat-related endonuclease 9 (CRISPR/Cas9) technology have achieved targeted interventions on endogenous genes in human CAR T cells. The CRISPR/Cas9 tool provides a simple method to generate multiple gene modifications. So far, 11 clinical trials for gene-edited CAR-T cells have been registered. CRISPR-edited CAR engineering enables immune cells to detect tumor-specific antigens and activate them after binding to their cognate antigens.

At Creative Biolabs, we have established a powerful platform that combines CAR cell therapy with CRISPR technology, which unleashes the therapeutic potential of CAR cell therapy and enables our global customers to push their projects into clinical trials. Our CRISPR/Cas9 system has been used to knock out and knock into multiple CAR cell genome editing sequences, such as CAR-T, CAR-NK, and CAR-MA cells, as well as CAR-stem cells.

Principles of CRISPR Gene-editing System

The use of CRISPR in genome editing is highly efficient and provides a simple and effective method to multiplex the processing of T cells. The CRISPR/Cas9 system has been used for sequence knockout and knock-in in mammalian genome editing (Figure 1). In principle, the deletion or insertion of the target gene is introduced by Cas9 nuclease guided by small RNA (sgRNA), which induces double-stranded DNA breaks, followed by non-homologous end joining (NHEJ) or homology-directed repair (HDR). In the NHEJ pathway, indels result in the deletion or insertion of nucleotides. In the HDR pathway, cofactors can promote genome recombination through two homology arms, leading to knock-in of the target gene.

Schematic of CRISPR gene-editing system.Fig.1 Schematic of CRISPR gene-editing system.

Applications of CRISPR Technology in Generation of Potent CAR T Cells

Currently, several CRISPR-based strategies are being used to develop next-generation CAR T cells through multiplexed genome editing. Such methods include knocking out endogenous genes (such as TCR, MHC, or autoantigens) to construct allogeneic universal CAR T cells, destroying inhibitory receptors (such as CTLA-4, PD-1, or LAG-3), and integration of the CAR cassette into the endogenous TCR α constant locus (TRAC) or CC chemokine receptor type 5 (CCR5) locus.

Gene-editing strategies for development of next generation CAR-T cell therapies.Fig.2 Gene-editing strategies for development of next generation CAR-T cell therapies.

Creative Biolabs provides a comprehensive list of CRISPR-edited jurkat cells, please browse the table below to explore more details.

KNOCK-OUT
TRAC TRBC PD-1 β2M CD7 LAG-3 CD4 CD5 CD52 Fas
CS-1 CTLA-4 PSCA MHC            
KNOCK-IN
TRAC exon 1 CCR5                

We also provide customized gene-editing services according to your interests and project needs. Please feel free to contact our team and get a complete solution about our CRISPR-edited CAR cell technology.

All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

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