Molecular switch, anti-HIV-1 chimeric antigen receptor having same, preparation method, NK cell, and application

A chimeric antigen receptor, HIV-1 technology, applied in the field of genes, can solve safety problems, toxic side effects and other problems, achieve the effects of high safety, increase proliferation and survival rate, and improve killing effect

Active Publication Date: 2019-01-25
SHANDONG XINRUI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, CAR-T cells will be overactive or out of control when infused into the body, resulting in certain toxic and side effects, which may cause safety issues in the application of HIV-1 treatment. In addition, CAR-T has a strong Personalized features make its industrialization a bottleneck in the application of HIV-1 treatment

Method used

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  • Molecular switch, anti-HIV-1 chimeric antigen receptor having same, preparation method, NK cell, and application
  • Molecular switch, anti-HIV-1 chimeric antigen receptor having same, preparation method, NK cell, and application
  • Molecular switch, anti-HIV-1 chimeric antigen receptor having same, preparation method, NK cell, and application

Examples

Experimental program
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Effect test

Embodiment 1

[0045] A molecular switch, the molecular switch is named RQR8, which has a nucleic acid sequence as described in SEQ ID NO.1. RQR8 is composed of signal peptide nucleic acid sequence (SEQ ID NO.2), CD20 mimic epitope nucleic acid sequence (SEQ ID NO.3), CD34 mimic epitope nucleic acid sequence (SEQ ID NO.4), CD20 mimic epitope The nucleic acid sequence (SEQ ID NO.3) and the CD8 hinge transmembrane region nucleic acid sequence (SEQ ID NO.5) are sequentially concatenated, and the complete nucleic acid sequence is shown in SEQ ID NO.1. Sangon Bioengineering (Shanghai) Co., Ltd. was commissioned to synthesize the nucleotide sequence.

Embodiment 2

[0047] The nucleic acid sequence of the anti-HIV-1 chimeric antigen receptor must contain the molecular switch RQR8 nucleic acid sequence. In this embodiment, the anti-HIV-1 chimeric antigen receptor comprises the CD4 antigen nucleic acid artificial sequence as described in SEQ ID NO.7 connected in sequence; the Linker nucleic acid artificial sequence as described in SEQ ID NO.8; 17b single-chain antibody nucleic acid artificial sequence as described in SEQ ID NO.9; CD8α hinge region nucleic acid artificial sequence as described in SEQ ID NO.10; NKG2D transmembrane region nucleic acid artificial sequence as described in SEQ ID NO.11; 2B4 co-stimulatory region nucleic acid artificial sequence as described in SEQ ID NO.12; CD3ζ signaling region nucleic acid artificial sequence as described in SEQ ID NO.13; T2A self-cleavage region nucleic acid artificial sequence as described in SEQ ID NO.14 ; RQR8 molecular switch region nucleic acid artificial sequence as described in SEQ ID N...

Embodiment 3

[0050] The chimeric antigen receptor preparation method comprises the following steps: (1) according to the artificial nucleic acid sequence of CD4 antigen, the artificial nucleic acid sequence of Linker, the artificial nucleic acid sequence of 17b single-chain antibody, the artificial nucleic acid sequence of CD8α hinge region, and the artificial nucleic acid sequence of NKG2D transmembrane region , 2B4 co-stimulatory region nucleic acid artificial sequence, CD3ζ signaling region nucleic acid artificial sequence, T2A self-cleavage region nucleic acid artificial sequence and RQR8 molecular switch region nucleic acid artificial sequence to synthesize the entire expression frame and insert it into the standard vector pUC to obtain pUC-CD4- 17bscFv-biCAR; (2) Carry out double digestion of pUC-CD4-17bscFv-biCAR, use agar gel electrophoresis to cut out the agar part of the CD4-17bscFv-biCAR DNA fragment, use the DNA extraction kit sol solution to treat, pass through the DF column and...

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Abstract

The invention provides a molecular switch, an anti-HIV-1 chimeric antigen receptor having the molecular switch, a preparation method, an NK cell, and an application. The molecular switch is named RQR8, which has a nucleic acid sequence shown in SEQ ID NO. 1. The chimeric antigen receptor provided by the invention is added with the molecular switch RQR8 for preventing and treating the adverse reaction of CD4-17bscFv-biCAR-NK, and the safety is improved.

Description

technical field [0001] The invention relates to the field of gene technology, in particular to a molecular switch, an anti-HIV-1 chimeric antigen receptor with the molecular switch, a preparation method, NK cells and applications. Background technique [0002] The clinical treatment of AIDS is mainly highly active antiretroviral therapy (Highly active antiretroviral therapy, HAART). The use of HAART has resulted in a mortality rate of almost 100% within five years and a survival rate of 91% within five years. Adults infected with HIV-1 in developed countries are estimated to live an average of more than 40 years from diagnosis. In developing countries, HIV-1-infected adults also have improved survival rates despite shorter life expectancies. Regardless of the remarkable achievements of the use of ART, we have to face a series of problems brought about by the use of ART. For example, ART has caused HIV-1 infection to develop into a chronic disease that is not easily cured. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N5/10C12N15/867A61K35/17A61P31/18
CPCA61K35/17A61P31/18C07K14/7051C07K16/1045C07K2317/76C07K2319/02C07K2319/03C07K2319/33C12N15/86C12N2740/15043
Inventor 刘明录王立新韩国英金海锋张传鹏冯建海韩庆梅刘敏王亮万磊
Owner SHANDONG XINRUI BIOTECH CO LTD
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