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Plasmid for silencing cytokine signal inhibitory factor 1 and expressing high mobility group B1 protein and tumor associated antigen and preparation method thereof

A technology for tumor-related antigens and cytokines, applied in the field of genetic engineering, can solve the problem of not exerting biological effects, and achieve the effects of stable and efficient transfection, no side effects, and good application prospects.

Inactive Publication Date: 2013-06-05
TIANJIN MEDICAL UNIV CANCER HOSPITAL
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AI Technical Summary

Problems solved by technology

The A-box protein can replace the full-length HMGB1 and bind to the corresponding receptor, but does not exert biological effects

Method used

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  • Plasmid for silencing cytokine signal inhibitory factor 1 and expressing high mobility group B1 protein and tumor associated antigen and preparation method thereof
  • Plasmid for silencing cytokine signal inhibitory factor 1 and expressing high mobility group B1 protein and tumor associated antigen and preparation method thereof
  • Plasmid for silencing cytokine signal inhibitory factor 1 and expressing high mobility group B1 protein and tumor associated antigen and preparation method thereof

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Embodiment Construction

[0050] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0051] The present invention simultaneously realizes the inhibition of DCs antigen presentation negative regulator SOCS1 through H1 and CMV double promoters, the activation of TLRs and the loading of DCs with tumor antigens, and detects its antigen presentation ability and activation effect on T cells by transfecting DCs in vitro , becoming a new and effective way to eliminate host-level self-tolerance, thereby inducing a more effective anti-tumor immune response.

[0052] In the constructed plasmid, the shRNA of SOCS1 was controlled by the human U6 RNA promoter, while the sequence encoding NY-ESO-1 and MAGE3 fusion protein was controlled by the CMV promoter. In this fusion protein, NY-ESO-1 is its full-length coding sequence, and MAGE-3 selects its HLA-A0201 (the HLA allele with the highest expression rate in Asian population) epitope p271-279 coding...

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Abstract

The invention discloses a plasmid for silencing cytokine signal inhibitory factor 1 and expressing high mobility group B1 protein and tumor associated antigen and a preparation method thereof, which belongs to genetic engineering technology. According to the invention, based on a gene recombination method, a cytokine signal inhibitory factor 1 small interfering RNA fragment shSOCS1, a fusion genecoding sequence of NY-ESO-1and MAGE3, and a coding sequence of HMGB1 are connected in the same vector of pIRES-EGFP; respective expression is guaranteed to be correct; and a plasmid coexpressing shSOCS1, HMGB1 and tumor associated antigen is constructed. A new-generation dendritic cell vaccine prepared by the plasmid of the invention can induce high-efficiency immune response of CTL cells with almost no side effects, and can become safe and effective immunotherapy.

Description

technical field [0001] The invention relates to genetic engineering technology, in particular to a plasmid for silencing cytokine signal inhibitor 1 and expressing high-mobility group protein B1 and tumor-associated antigen and a preparation method thereof. technical background [0002] Suppressor of cytokine signaling 1 (SOCS1) is a newly discovered inhibitor of antigen presentation, which has been confirmed to play a key role in maintaining self-tolerance and limiting anti-tumor immunity. SOCS1 is a key negative regulator of the signaling of various proinflammatory cytokines in T cells and antigen-presenting cells, including IFN-g, IL-2, IL-6, IL-7, IL-12, IL -15 etc. As a feedback mechanism, SOCS1 is induced after proinflammatory response stimulation, which can act as a false substrate to prevent the binding of JAK and STAT, and at the same time increase the ubiquitination and degradation of JAK. In addition, SOCS1 can also inhibit NF-κB signaling pathway to negatively ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/66
Inventor 任秀宝魏枫
Owner TIANJIN MEDICAL UNIV CANCER HOSPITAL
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