Recombinant CAR19-IL24 gene, lentiviral vector, CAR19-IL24-T cell and application of recombinant CAR19-IL24 gene, lentiviral vector and CAR19-IL24-T cell

A technology of CAR19-IL24 and lentiviral vector, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, receptors/cell surface antigens/cell surface determinants, etc., to achieve high specificity, good therapeutic effect, and strong tumor killing function Effect

Active Publication Date: 2019-05-10
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently no CAR-T cell products that autocrine IL-24

Method used

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  • Recombinant CAR19-IL24 gene, lentiviral vector, CAR19-IL24-T cell and application of recombinant CAR19-IL24 gene, lentiviral vector and CAR19-IL24-T cell
  • Recombinant CAR19-IL24 gene, lentiviral vector, CAR19-IL24-T cell and application of recombinant CAR19-IL24 gene, lentiviral vector and CAR19-IL24-T cell
  • Recombinant CAR19-IL24 gene, lentiviral vector, CAR19-IL24-T cell and application of recombinant CAR19-IL24 gene, lentiviral vector and CAR19-IL24-T cell

Examples

Experimental program
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Embodiment Construction

[0035] 1 Construction of CAR19 expression cassette and lentiviral vector

[0036](1) Find the FMC63scFv sequence that specifically recognizes the CD19 molecule according to existing reports and bioinformatics methods, design the transmembrane region and intracellular signal transduction region sequence and gene synthesis sequence (CAR19), and design Bbs1 enzyme at both ends of the CAR19 sequence Cut site, the sequence loaded in the pUC57 plasmid (pUC57-CAR19).

[0037] (2) Bbs1 digestion of pUC57-CAR19 will generate BamH1 and Age1 cohesive ends at the 5' and 3' ends of the CAR19 sequence, respectively. At the same time, the second-generation lentiviral vector plasmid pHQ-009 was digested with BamH1 and Age1. (with enzyme digestion system)

[0038] (3) Gel recovery pHQ-009 backbone 7379bp and CAR19 sequence 1534bp. Use T4 DNA Ligase to ligate to construct plasmid pHQ-010, and the ligation time should not exceed 10 minutes at room temperature (25°C) or overnight in a water ba...

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Abstract

The invention discloses a recombinant CAR19-IL24 gene, a lentiviral vector, a CAR19-IL24-T cell and application of the recombinant CAR19-IL24 gene, lentiviral vector and CAR19-IL24-T cell.

Description

technical field [0001] The invention belongs to the technical field of tumor cell immunity, and specifically relates to a recombinant CAR19-IL24 gene, a lentiviral vector, CAR19-IL24-T cells and applications thereof. Background technique [0002] Chimeric Antigen Receptor T-Cell Immunotherapy (Chimeric Antigen Receptor T-Cell Immunotherapy), also known as CAR-T cell immunotherapy, is a new type of immunotherapy that uses chimeric antigen receptor T cells for immunotherapy. Chimeric antigen receptor T cells (CAR-T) are T cells that have been modified to express chimeric antigen receptor (CAR) molecules on their surface. The CAR molecule expressed on the T cell membrane will "redirect" the T cell, endowing it with the ability to specifically recognize and kill the target cells expressing the CAR molecular target, thereby specifically eliminating the tumor target cells in the patient. A chimeric antigen receptor (CAR) generally includes the antigen-binding portion of an antibo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/867C12N5/10
CPCA61K39/0011A61K2039/5156A61K2039/5158C07K14/7051C07K2319/03
Inventor 梁德生刘雄昊胡乾邬玲仟刘慕君
Owner CENT SOUTH UNIV
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