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Application of TSPO (Translocator Protein) in treatment of brain glioma and recombinant herpes simplex virus as well as preparation method and application of recombinant herpes simplex virus

A herpes simplex virus and glioma technology, applied in the field of genetic engineering, can solve the problems that the effectiveness of the HSV-1 oncolytic virus cannot meet the treatment needs, the number and replication ability of the oncolytic virus are low, and the efficacy of the oncolytic virus is low. , to achieve good application prospects, improve killing efficiency, and reduce side effects

Active Publication Date: 2018-05-08
BEIJING NEUROSURGICAL INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the effectiveness of HSV-1 oncolytic virus still cannot meet the treatment needs
Reasons for the low efficacy of HSV-1 oncolytic viruses include the rapid growth of gliomas leading to low relative numbers and replication capacity of oncolytic viruses

Method used

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  • Application of TSPO (Translocator Protein) in treatment of brain glioma and recombinant herpes simplex virus as well as preparation method and application of recombinant herpes simplex virus
  • Application of TSPO (Translocator Protein) in treatment of brain glioma and recombinant herpes simplex virus as well as preparation method and application of recombinant herpes simplex virus
  • Application of TSPO (Translocator Protein) in treatment of brain glioma and recombinant herpes simplex virus as well as preparation method and application of recombinant herpes simplex virus

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Embodiment approach

[0028] According to a preferred embodiment of the present invention, the vector is a herpes simplex virus that has deleted the genes encoding ICP34.5 and ICP47, and the insertion site of the foreign nucleotide sequence is the deletion of the genes encoding ICP34.5 on the vector. and / or the location of the ICP47 gene.

[0029] According to the present invention, in order to effectively independently translate and express the foreign nucleotide sequence, the foreign nucleotide sequence of the present invention preferably includes a promoter and a termination sequence, and the foreign nucleotide sequence preferably includes A promoter, an optional start codon and a termination sequence (which may be a stop codon), and an optional linker sequence and / or a PolyA sequence, in such preferred cases, when the recombinant herpes simplex virus infects a host cell and When expressing its own gene, it can transcribe an independent and complete target mRNA fragment.

[0030] In another pre...

Embodiment 1

[0059] This example is used to illustrate the construction of the recombinant herpes simplex virus provided by the present invention.

[0060]According to application number 2004100064921, the ICP34.5 gene and the ICP47 gene knockout of wild-type HSV-1 virus (the GenBank number of its gene sequence: NC_001806, the same below) are knocked out by the method described in the patent application of authorized announcement number CN1283803C, and in HSV The ICP34.5 gene knockout of -1 virus is inserted into an exogenous nucleotide sequence, but the difference is that Vero cells are used as host cells in this embodiment. The exogenous nucleotide sequence includes: U6 promoter, shTSPO#2, termination sequence (TTTTTT) from the 5' end to the 3' end. Sequencing at Suzhou Jinweizhi Company confirmed that the foreign nucleotide sequence was correctly inserted into the herpes simplex virus vector to obtain the recombinant virus vector. Successfully constructed recombinant viral vectors were...

Embodiment 2

[0062] This example is used to illustrate the construction of the recombinant herpes simplex virus provided by the present invention.

[0063] According to the application number 2004100064921, the method recorded in the patent application of the authorized announcement number CN1283803C will knock out the ICP34.5 gene and the ICP47 gene of the wild-type HSV-1 virus, and, at the position of knocking out the ICP34.5 gene of the HSV-1 virus Insert the artificial chemically synthesized exogenous nucleotide sequence 1, and insert the artificially chemically synthesized exogenous nucleotide sequence 2 at the position of the knockout ICP47 gene of the HSV-1 virus. The difference is that this embodiment uses Vero cells as the host cell. Wherein, the exogenous nucleotide sequence 1 includes: U6 promoter, shTSPO#2, and termination sequence (TTTTTT) from the 5' end to the 3' end. The exogenous nucleotide sequence 2 includes in sequence from the 5' end to the 3' end: CMV promoter, gene ...

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Abstract

The invention relates to the field of gene engineering, and discloses a recombinant herpes simplex virus as well as a preparation method and application of the recombinant herpes simplex virus. Particularly, the recombinant herpes simplex virus comprises a carrier and an exogenous nucleotide sequence, wherein the carrier is a herpes simplex virus of at least one of a gene lacking codes ICP34.5 andICP47 and a selected gene lacking the codes ICP6, TK and UNG; an insertion site of the exogenous nucleotide sequence is at least one of positions of the genes lacking the codes ICP34.5, ICP47, ICP6,TK and UNG on the carrier; the exogenous nucleotide sequence comprises a nucleotide sequence capable of reducing the expression level of the TSPO gene. The invention also discloses application of reduction of the expression level of the TSPO gene in the brain glioma cell in preparation of medicine for treating the brain glioma. The recombinant herpes simplex virus disclosed by the invention has agood anti-brain glioma effect.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the application of reducing the expression level of TSPO gene in glioma cells in the preparation of medicines for treating glioma, as well as recombinant herpes simplex virus and its preparation method and application. Background technique [0002] Glioma is the most common tumor of the central nervous system. The World Health Organization (WHO) divides glioma into four grades from I to IV. I and II are low-grade gliomas. High-grade gliomas, in which the high-grade gliomas are more aggressive, the prognosis of patients is worse, and the median survival time is only 18 months (Ⅲ) and 12 months (Ⅳ). The treatment of glioma usually adopts surgical resection, supplemented by comprehensive treatment such as radiotherapy and chemotherapy; however, it is difficult to completely remove glioma with surgery, the recurrence rate is high, and it is very easy to develop tolerance to radioth...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K35/763A61P35/00C12R1/93
CPCA61K35/763C12N7/00C12N2710/16621C12N2710/16632
Inventor 刘福生李小鹏田超张俊文
Owner BEIJING NEUROSURGICAL INST
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