A dual-gene targeted therapy system for recurrent HSK

A targeted therapy and dual-gene technology, applied in gene therapy, genetic engineering, plant gene improvement, etc., can solve the problems of keratitis that is easy to recur and difficult to cure, and achieves the solution of latent infection, high-efficiency treatment effect, and suppression of latent virus infection Effect

Active Publication Date: 2018-11-30
陕西省眼科研究所
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  • Abstract
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Problems solved by technology

[0003] The purpose of the present invention is to provide a dual-gene targeted therapy system for the treatment of recurrent HSK to solve the problem that herpes simplex virus keratitis is easy to relapse and difficult to cure

Method used

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  • A dual-gene targeted therapy system for recurrent HSK
  • A dual-gene targeted therapy system for recurrent HSK
  • A dual-gene targeted therapy system for recurrent HSK

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Embodiment Construction

[0031] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0032] The invention discloses a dual-gene targeted therapy system rdHSV1-IFNγ-siRNA, such as figure 2 , image 3 As shown, the construction method of the dual-gene targeted therapy system rdHSV1-IFNγ-siRNA is as follows:

[0033] Delete the ICP4, ICP34.5 and ICP27 genes in the herpes simplex virus HSV1 to make it unable to replicate, mutate the VP16 gene, and transform it into an HSV 1764 4- / 27+ / RL1+ skeleton virus, such as figure 1 As indicated, it was made into a replication-deficient vector rdHSV1, wherein the herpes simplex virus HSV1 was purchased from Beijing Nuosai Genome Research Center Co., Ltd.

[0034] Isolate human blood lymphocytes (purchased from Sangon Bioengineering (Shanghai) Co., Ltd.), inoculate HSV1 virus to induce IFNγ gene, extract total lymphocyte RNA, reverse cDNA, and obtain IFNγ gene sequence. The sequence is spe...

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Abstract

The invention discloses a constructing method of a double-gene targeting treatment system rdHSV1-IFNgamma-siRNA. The constructing method specifically comprises the following steps of (1) deleting genes ICP4, ICP34.5 and ICP27 in herpes simplex virus HSV1, and mutating a gene VP16, so as to obtain a replication-defective type carrier rdHSV1; screening a target sequence of a gene LAT, proofing the target site, and forming an siRNA nucleotide sequence which is shown in SEQ ID NO:2 in a sequence list; respectively connecting a sequence of a gene IFNgamma and the siRNA sequence into a shuttle plasmid, and performing homologous recombination with the replication-defective type carrier rdHSV1, so as to obtain the double-gene targeting treatment system rdHSV1-IFNgamma-siRNA. The constructing method has the advantages that the problems of virus latent infection and recurrence in the existing HSK are solved; the existing most advanced HSV virus carrier is used as a tool to establish a double-gene targeting treatment method for the latent marker gene LAT of the HSV virus and the virus activating, and the virus latent part is specifically positioned to perform gene silencing and intervention treatment.

Description

【Technical field】 [0001] The invention belongs to the field of biotechnology, and in particular relates to a dual-gene targeted therapy system for treating recurrent HSK. 【Background technique】 [0002] Herpes simplex keratitis (Herpes Simplex Keratitis, HSK) is the most common viral keratitis, and its morbidity and blindness rate rank first among corneal diseases. The disease is prone to relapse due to the latent infection of Herpes Simplex Virus (HSV) in human trigeminal ganglion and corneal tissue. Severe cases are protracted and eventually blind. Herpes simplex virus (HSV) is highly infectious to humans. Among adults over the age of 20, the positive rate of serum antibodies reaches 90%. The primary infection is only seen in children who have no immunity to the virus, mostly children aged 6 months to 5 years old. After the primary infection, the virus stays latent in the body for a lifetime and waits for recurrence. Secondary infection is more common in children over 5...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/869C12N15/23C12N15/113A61K48/00A61K31/7088A61K47/46A61P27/02A61P31/22
Inventor 徐琨刘先宁朱秀萍程燕郑璇
Owner 陕西省眼科研究所
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