A method for rapid preparation of pd-1ˉt cells by efficient gene editing and its application

A PD-1 and gene editing technology, applied in the field of biomedicine, can solve the problem of low plasmid transfection efficiency PD-1 gene editing efficiency, etc., and achieve the effect of improving gene editing efficiency, high efficiency and strong anti-tumor effect.

Active Publication Date: 2021-08-31
GUANGXI MEDICAL UNIVERSITY
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Problems solved by technology

[0005] However, although the existing CRISPR / Cas9 gene technology has been applied in many fields, it has also shown good results, but in terms of tumor treatment, especially knocking out the PD-1 gene on T cells is not in the public technology. Not involved, in addition, the existing CRISPR / Cas9 knockout PD-1 gene technology did not find the combined application of high penetration peptide small molecule TAT, so there are problems of low plasmid transfection efficiency and PD-1 gene editing efficiency

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  • A method for rapid preparation of pd-1ˉt cells by efficient gene editing and its application
  • A method for rapid preparation of pd-1ˉt cells by efficient gene editing and its application
  • A method for rapid preparation of pd-1ˉt cells by efficient gene editing and its application

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Embodiment Construction

[0041] The present invention will be further described in detail below in conjunction with specific embodiments, and the given examples are only for clarifying the present invention, not for limiting the scope of the present invention.

[0042] The experimental methods in the following examples are conventional methods unless otherwise specified.

[0043] Materials, reagents, etc. used in the following examples. Unless otherwise specified, all can be obtained from commercial sources.

[0044] One, the material and reagent used in the embodiment:

[0045] Peripheral blood was collected from healthy volunteers, and the lymphocyte separation liquid was purchased from Tianjin Haoyang Biological Products Company.

[0046] Human liver cancer HepG2 cells (ATCC, product number HB-8065), the public can obtain this application material from the applicant, which is only used for repeating the experiment of the present invention, and cannot be used for other purposes.

[0047] 2. High-...

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Abstract

The invention discloses a method for obtaining PD-1-T cells, comprising the following steps: (1) obtaining human peripheral blood T lymphocytes; (2) plasmid transfection; (3) preparation of fusion cells; (4) PD Expansion of ‑1‑T tumor-specific T cells. The invention also discloses the application of the method for obtaining PD-1-T cells. The PD-1-T cells obtained by the method of the invention exclude negative inhibitory regulatory receptors and have a good anti-tumor effect. The adoptively treated cells effectively prolong the survival time of tumor-bearing mice and are suitable for promotion in the medical field.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for rapidly preparing PD-1ˉT cells by efficient gene editing and its application. Background technique [0002] T cells in the body's immune system are the main component of lymphocytes. T lymphocytes are mainly used to resist foreign invaders and have various biological functions. It is the body's main "warrior" against disease invasion, infection and tumor formation. At present, the methods of tumor treatment are generally surgery, radiotherapy and chemotherapy. There are some problems in these methods, among which immune escape phenomenon will appear in adoptive therapy, which will affect the anti-tumor effect. [0003] PD-1 (programmed death1), that is, programmed death receptor 1, is a negative inhibitory regulatory receptor. When T cells are activated, the expression of PD-1 can be significantly increased. Through the combined effect with the ligand PD-L1, i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/85A61K35/17A61P35/00
CPCA61K35/17C07K14/70521C12N5/0636C12N15/85C12N2510/00C12N2800/80C12N2810/10
Inventor 赵永祥周素芳李桂银
Owner GUANGXI MEDICAL UNIVERSITY
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