Chimeric antigen receptor gene engineering vector and immune cells, and applications thereof

A chimeric antigen receptor and genetic engineering technology, applied in genetic engineering, blood/immune system cells, applications, etc., can solve the problems of many side effects and poor specificity of treatment, and achieve the goal of ensuring safety and reducing the probability of virus recombination Effect

Pending Publication Date: 2019-08-16
SHANGHAI CHILDRENS MEDICAL CENT AFFILIATED TO SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In summary, the current first-, second- and third-generation chimeric

Method used

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  • Chimeric antigen receptor gene engineering vector and immune cells, and applications thereof
  • Chimeric antigen receptor gene engineering vector and immune cells, and applications thereof
  • Chimeric antigen receptor gene engineering vector and immune cells, and applications thereof

Examples

Experimental program
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Embodiment 1

[0058] Example 1: 1) First, construct a chimeric antigen receptor genetic engineering vector. The corresponding sequence was synthesized by gene synthesis method and verified by sequencing. Select a suitable restriction site to insert the above sequence into a suitable position in the adenovirus, retrovirus or lentivirus vector, and verify it by sequencing. Such as figure 1 and figure 2 As shown, chimeric antigen receptor genetic engineering vectors include:

[0059] The RRE sequence is inserted into the upstream of the multiple cloning site of the vector, the WPRE sequence and cPPT sequence are inserted into the downstream of the multiple cloning site, and the EF1A promoter is inserted between the RRE sequence and the upstream of the multiple cloning site to initiate the insertion of multiple cloning chimeric antigen receptor gene. The sequence upstream of the chimeric antigen receptor structural protein is optimized, and the optimized sequence is shown in SEQID NO60, so...

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Abstract

The invention belongs to the field of cell therapy, and discloses a chimeric antigen receptor gene engineering vector and immune cells, and applications thereof, wherein the chimeric antigen receptorgene engineering vector comprises a lentivirus vector skeleton, regulatory elements connected onto the lentivirus vector skeleton and a chimeric antigen receptor gene sequence. The regulatory elementsinclude a WPRE element, a cPPT element and an RRE element. The chimeric antigen receptor gene sequence comprises a costimulatory signal pathway sequence and an effector signal pathway sequence whichare independently expressed; or the chimeric antigen receptor gene sequence comprises a costimulatory signal pathway sequence and an effector signal pathway sequence which are in fused expression. Thechimeric antigen receptor gene engineering vector has the advantages of good safety, high transfection efficiency, rapid amplification of transfected immune cells, and great improvement of killing effect. In addition, the recognition on target cells by the chimeric antigen receptor immune cells is more precise than the recognition on target cells by chimeric antigen receptor immune cells with single target, and the off-target effect of single target can be prevented.

Description

technical field [0001] The invention belongs to the field of cell therapy, and in particular relates to a chimeric antigen receptor genetic engineering carrier, immune cells and applications thereof. The genetic engineering vector can be stably transfected into autologous or allogeneic immune cells after virus packaging, and can specifically kill target cells with target molecules in the body through dual-target recognition, thereby playing a clinical therapeutic role. Background technique [0002] Chimeric antigen receptor-Chimeric antigen receptor, abbreviated as CAR. Among them, chimeric antigen receptor cell therapy based on T cells is referred to as CART therapy, chimeric antigen receptor cell therapy based on NK cells is referred to as CAR-NK therapy, and there is also chimeric antigen receptor cell therapy based on γ / δT cells, called For Universal-CAR. No matter what kind of cells are used, the strong killing ability of corresponding immune cells on target cells is ...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10A61K35/17A61P35/00A61P35/02A61P37/00A61P31/12A61P31/10A61P31/04
CPCC12N15/86C12N5/0636C12N5/0646A61K35/17A61P35/00A61P35/02A61P37/00A61P31/12A61P31/10A61P31/04C12N2740/15043C12N2510/00C12N5/10C12N15/867Y02A50/30
Inventor 李本尚
Owner SHANGHAI CHILDRENS MEDICAL CENT AFFILIATED TO SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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