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Chimeric antigen receptor and expression gene thereof, optically-controlled and regulated T-cell modified by chimeric antigen receptor and application thereof.

A chimeric antigen receptor and gene expression technology, applied in genetically modified cells, cells modified by introducing foreign genetic material, receptors/cell surface antigens/cell surface determinants, etc. Sexual problems, etc.

Active Publication Date: 2017-12-08
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 expression gene thereof, optically-controlled and regulated T-cell modified by chimeric antigen receptor and application thereof.
  • Chimeric antigen receptor and expression gene thereof, optically-controlled and regulated T-cell modified by chimeric antigen receptor and application thereof.
  • Chimeric antigen receptor and expression gene thereof, optically-controlled and regulated T-cell modified by chimeric antigen receptor and application thereof.

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Experimental program
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preparation example Construction

[0119] See image 3 In one embodiment, a method for preparing light-controlled chimeric antigen receptor modified T cells includes the following steps S110 to S130.

[0120] S110. Provide chimeric antigen receptor expression genes, which are used to encode chimeric antigen receptors, wherein the chimeric antigen receptors include a first fusion protein and a second fusion protein, and the first fusion protein includes The antigen binding region, hinge protein, first transmembrane protein and Arabidopsis cryptochrome protein-2 are connected in sequence, and the second fusion protein includes the second transmembrane protein, basic helix loop helix protein-1 and cell Internal signaling protein.

[0121] Specifically, for the structure of the chimeric antigen receptor (CAR), please refer to the above description, and will not be repeated here.

[0122] In one embodiment, the expression of the encoded first fusion protein is anti-mesothelin-CD8αhinge-CD28TM-CD28-4-1BB-CRY2. The express...

Embodiment 1

[0136] Preparation of Light-Controlled Chimeric Antigen Receptor Modified T Cells

[0137] The nucleic acid sequence encoding the first fusion protein (including the nucleotide sequence shown in SEQ ID No.1, the nucleotide sequence shown in SEQ ID No.2, and the nucleotide sequence shown in SEQ ID No.3 are sequentially connected 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] The nucleic acid sequence encoding the second fusion protein (including 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 which are sequentially connected 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 the nucleic acid sequence encoding the second fusion protein are connected by a nucleic acid seq...

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

Claims

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Application Information

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Patent Type & Authority Applications(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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