CRISPR-Cas9 for Knocking Out Negative Immune Checkpoint Genes

CRISPR-Cas9 in tumor immunotherapy

Gene editing technology is an emerging molecular biotechnology that artificially changes the specific gene locus in a certain nucleotide sequence to change its expression traits through inserting, removing, replacing or modifying the specific target gene in the genome. As an emerging gene editing technology, CRISPR-Cas9 has been widely applied in the field of tumor therapy via targeting knockout of tumor immune checkpoint molecules or rapid and simple gene editing, that has significantly reduced the operational difficulty of tumor immunotherapy and greatly promoted the development of tumor immunotherapy research.

CRISPR-Cas9 in immune checkpoint targets

As well-known immune checkpoints, PD-1 and CTLA-4 can significantly inhibit the activity of T cells, which in turn make tumor cells easy to escape the immune mechanism of the human body, leading to poor therapeutic effect of T cells. At present, blocking antibodies with immune checkpoint inhibitors is commonly used as an effective method to suppress the immune response of T cells by blocking immune regulators.

CRISPR-Cas9 gene editing technology is a novel cancer immunotherapy method. Several research papers published have indicated that CRISPR-Cas9 gene editing technology can achieve the similar anti-tumor effect with immune checkpoint inhibitors by knocking out genes involved in negative immune regulation. Cas9 and sgRNA are introduced into T cells by electroporation method and PD-1 gene will be knocked out to reduce its expression. No effects of PD-1 on the activation of T cells was found during long-term in vitro culture, therefore, it can be concluded that T cells can better exert their anti-tumor effect after PD-1 gene knockout. This new blocking therapy is often used in combination with adoptive T cell immunotherapy to enhance the activity of immune cells.

Figure 1. Generating PD-1 KO antigen-specific polyclonal CTL lines by CRISPR-Cas9 system.

Our Platform

Creative BioMart offers CRISPR-Cas9 CRO platform for knocking out negative immune checkpoint genes such as PD-1 and CTLA-4. We will discuss with research teams of customers and make the best plan for immune checkpoint genes knocking out. We are devoted to provide mature platform for academic and industrial researchers who are focused on tumor immunotherapy.

Strengths

• Experienced scientists on gene editing and immune-oncology areas.
• Matured CRISPR-Cas9 platform for gRNA design and CRISPR-gRNA plasmid construction.
• CRISPR gRNA library.
• Off-the-shelf inhibitory immune checkpoint antibodies library.

References

1. Shu Su et al. (2016) CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients. Scientific Reports 6: 20070.
2. Chi Zhang et al. (2018) Genetic abrogation of immune checkpoints in antigen-specific cytotoxic T-lymphocyte as a potential alternative to blockade immunotherapy. Scientific Reports 8: 5549.
3. Wanghong Hu et al. (2019) CRISPR/Cas9-mediated PD-1 disruption enhances human mesothelin-targeted CAR T cell effector functions. Cancer Immunology, Immunotherapy 68-3:365–377.