Chimeric Antigen Receptor and Its Expression Gene, Light Controlled Modulation of Chimeric Antigen Receptor Modified T Cell and Its Application

A technology of chimeric antigen receptors and gene expression, applied to genetically modified cells, cells modified by introducing foreign genetic material, receptors/cell surface antigens/cell surface determinants, etc., can solve poor specificity and side effects of treatment Waiting for questions

Active Publication Date: 2020-12-25
深圳华云生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In summary, traditional chimeric antigen receptors have many side effects and poor specificity.

Method used

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  • Chimeric Antigen Receptor and Its Expression Gene, Light Controlled Modulation of Chimeric Antigen Receptor Modified T Cell and Its Application
  • Chimeric Antigen Receptor and Its Expression Gene, Light Controlled Modulation of Chimeric Antigen Receptor Modified T Cell and Its Application
  • Chimeric Antigen Receptor and Its Expression Gene, Light Controlled Modulation of Chimeric Antigen Receptor Modified T Cell and Its Application

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preparation example Construction

[0119] see image 3 , a method for preparing light-regulated chimeric antigen receptor-modified T cells according to one embodiment, comprising the following steps S110-S130.

[0120] S110, providing a chimeric antigen receptor expression gene, the chimeric antigen receptor expression gene is used to encode a chimeric antigen receptor, wherein the chimeric antigen receptor includes a first fusion protein and a second fusion protein, and the first fusion protein includes The antigen-binding domain, hinge protein, first transmembrane protein and Arabidopsis cryptochrome protein-2 are connected sequentially, and the second fusion protein includes the second transmembrane protein, basic helix loop helix protein-1 and cytoplasmic protein-1 connected sequentially. Internal signaling proteins.

[0121] Specifically, the structure of the chimeric antigen receptor (CAR) can be referred to above, and will not be repeated here.

[0122] In one embodiment, the expression of the encoded ...

Embodiment 1

[0136] Preparation of light-regulated chimeric antigen receptor-modified T cells

[0137] Synthesize the nucleic acid sequence (comprising the nucleotide sequence shown in SEQ ID No.1, the nucleotide sequence shown in SEQ ID No.2, the nucleotide sequence shown in SEQ ID No.3 connected sequentially by the gene company) of the first fusion protein respectively The nucleotide sequence and the nucleotide sequence shown in SEQ ID No.4). The expression of the first fusion protein is anti-mesothelin-CD8αhinge-CD28TM-CD28-4-1BB-CRY2.

[0138] Synthesize the nucleic acid sequence (comprising the nucleotide sequence shown in SEQ ID No.5, the nucleotide sequence shown in SEQ ID No.6 and the nucleotide sequence shown in SEQ ID No.7 connected sequentially by the gene company respectively) of encoding the second fusion protein nucleotide sequence). The expression of the second fusion protein is CD28-4-1BB-CIB1-CD3ζ.

[0139] The nucleic acid sequence encoding the first fusion protein and...

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Abstract

The invention relates to a chimeric antigen receptor and an expression gene thereof, optically-controlled and regulated T-cell modified by the chimeric antigen receptor and application thereof. The expression gene of the chimeric antigen receptor comprises a first fusion protein expression gene and a second fusion protein expression gene, wherein the first fusion protein expression gene comprises an antigen bonding area expression gene, a hinge protein expression gene, a first transmembrane protein expression gene, and an arabidopsis cryptochrome-2 expression gene which are sequentially connected; the second fusion protein expression gene comprises a second transmembrane protein expression gene, a basic helix-loop-helix-1 expression gene and an intracellular signaling protein expression gene which are sequentially connected. The innovatively designed expression gene of the chimeric antigen receptor has the advantages that the expression gene is successfully imported into the T cell to form the T cell modified by the chimeric antigen receptor, the immunoreaction is only produced under the particular blue light condition and particular antigen condition, and the immunoreaction of the chimeric antigen receptor is controllable; the side effect in therapy is low, and the specificity is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a chimeric antigen receptor and its expression gene, light-controlled regulated chimeric antigen receptor-modified T cells and applications thereof. Background technique [0002] Chimeric antigen receptor (CAR) is an artificially constructed transmembrane molecule encoded by a fusion gene. It generally consists of an extracellular antigen-binding domain, a transmembrane domain and an intracellular signaling domain. CAR-T, that is, chimeric antigen receptor (CAR) modified T cells. The application of CAR-T cells in cancer treatment has shown excellent efficacy and great potential. For example, Novartis announced the phase 2 clinical data of its CD19-targeting CAR-T (CTL019) in the treatment of refractory and relapsed acute lymphoblastic leukemia (ALL) at the ASH conference on December 5, 2015. Phase 2 data Similar to the phase 1 data published in 2014, the complete remission rate (C...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62C12N5/10C07K19/00A61K35/17A61P31/14A61P31/20A61P35/00
CPCA61K35/17C07K14/7051C07K16/081C07K16/10C07K16/30C07K2319/02C07K2319/03C07K2319/33C12N2510/00
Inventor 黄彪陈雪梅刘韬
Owner 深圳华云生物技术有限公司
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