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Modular CAR-T Cell Generation with AAV-Cpf1

At Creative Biolabs, we are focused on the cutting-edge technologies, and our scientists are passionate about providing innovative and valuable solutions to advance the CAR-T projects of our clients. To enable flexible and efficient CAR-T cell generation, we have developed the most cutting-edge platform - trans-activating CRISR (clustered regularly interspaced short palindromic repeats) RNA (tracrRNA)-independent CRISPR-Cpf1 system with adeno-associated virus (AAV) to generate double-knock-in CAR-T cells or homology-directed-repair knock-in and immune-checkpoint knockout CAR-T cells at high efficiency in one step.

Our Inspiration

Recently, one-step generation of modular CAR-T cells with AAV-Cpf1 was published in Nature Methods journal, and our scientists are excited about this innovative approach. By leveraging trans-activating CRISPR (clustered regularly interspaced short palindromic repeats) RNA (tracrRNA)-independent CRISPR-Cpf1 systems with adeno-associated virus (AAV), a stable CAR-T cell with homology-directed-repair knock-in and immune-checkpoint knockout was built at very high efficiency by streamlined genome engineering (see Fig.2). If you are interested in CRISPR-Cas9-based TRAC-CAR-T cells, please visit here to learn more. Compared with CRISPR-Cas9 based approach, CRISPR-Cas12a/Cpf1 is independent of RNaseIII and tracrRNA (see Fig.1). This enables the modular CAR-T cell generation via T cell engineering with simplicity and precision.

Differences between Cas9 and Cpf1.

Fig.1 Differences between Cas9 and Cpf1.

Modular CAR-T Cell Generation with AAV-Cpf1

As a full-service CRO, we are inspired by this method and our scientists try their best to develop this platform at the earliest convenience. We have established this platform to advance your CAR-T cells more efficient. Knock-in and knock-out gene editing can be realized at flexible combinations. Modular bispecific knock-in CAR-T cells can be made stable. Before delivery modular CAR-T cells to our clients, validation assays can be conducted as per our clients’ requirements. With this approach, modular CAR-T cells can be generated more flexible and highly efficient. Compared with popular Cas-based methods, the AAV-Cpf1 system can generate double-knock-in and knock-in/knock-out CAR-T cells more efficiently. Functional assays (cytokine production and cancer killing) of AAV-Cpf1 CAR-T cells have been validated, and lower levels of exhaustion markers were expressed. This method is exciting and it opens new capabilities of T cell engineering with simplicity and precision.

Schematic of LbCpf1 mRNA electroporation combined with AAV-delivered crRNA and HDR template (AAV-Cpf1).

Fig.2 Schematic of LbCpf1 mRNA electroporation combined with AAV-delivered crRNA and HDR template (AAV-Cpf1). (Xiaoyun, 2019)

Highlight Features

This platform had been validated by our scientists via cytokine production and cancer cell killing assays. We are dedicated to providing the most valuable solutions to our clients with consistent performance.

  • Multiple gene editing including knock-in and knock-out,
  • Flexible and highly efficient approach,
  • Stable CAR-T cell with low levels of exhaustion markers,
  • With extensive experience of CAR-T projects.

Creative Biolabs is dedicated to advancing your CAR-T projects with versatile systems. Our innovative and collaborative approach enables us to provide a broader range of services proximate to our clients. We are committed to trying our best to provide CAR-T services. If you are interested in any CAR related technologies, please feel free to contact us to learn more.

Reference

  1. Dai, Xiaoyun, et al. "One-step generation of modular CAR-T cells with AAV-Cpf1." Nature methods (2019): 1.

Online Inquiry

For any technical issues or products/services related questions, please leave your contacts as below, and our team will contact you at earliest convenience to let you know how we can be involved in your projects.

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