A method for efficient knockout of cd96 gene in nk cells

A high-efficiency technology for NK cells, applied in the field of gene editing, to achieve high knockout efficiency, improved recognition and killing activity, obvious application prospects and clinical application value
CN112175905BActive Publication Date: 2022-07-22SHANGHAI UNIV OF T C M
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
SHANGHAI UNIV OF T C M
Publication Date
2022-07-22

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Abstract

The invention discloses a method for efficiently knocking out the CD96 gene in NK cells. The method uses a CRISPR / Cas9 gene editing system to introduce a complex of sgRNA and Cas9 protein into amplified NK cells by electroporation transfection wherein: the sequence of the sgRNA is selected from any one of the sequences shown in SEQ ID NO.1-10. The method of the present invention can realize the knockout of CD96 gene in NK cells, not only the knockout efficiency is high, but also the obtained NK cells that do not express CD96 can relieve the immunosuppressive effect of CD155+ tumor cells on NK cells. Wild-type NK cells show stronger anti-tumor activity in both in vivo and in vitro experiments, significantly improving the recognition and killing activities of NK cells on tumor cells, and are expected to be developed into safe and effective anti-tumor biological agents.
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Description

technical field

[0001] The invention relates to a method for efficiently knocking out CD96 gene in NK cells, and belongs to the technical field of gene editing. Background technique

[0002] Natural killer cells (NK cells) are important immune cells in the body. They are derived from bone marrow lymphoid stem cells. Their differentiation and development depend on the bone marrow and thymus microenvironment. They are mainly distributed in bone marrow, peripheral blood, liver, spleen and lung. and lymph nodes. Different from T and B cells, NK cells are a type of lymphocytes that can non-specifically kill tumor cells and virus-infected cells without prior sensitization. It is not only related to anti-tumor, anti-viral infection and immune regulation, but also in some cases. It is involved in the occurrence of hypersensitivity reactions and autoimmune diseases, and can recognize target cells and killing mediators.

[0003] Studies have shown that although NK cells have powerfu...

Claims

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