Anti-HIV/AIDS gene targeting vector, construction method and application

Abstract

The invention belongs to the field of bioengineering and particularly relates to construction and acquisition of a targeting vector for inserting a control type ZFN expression element with a functionof targeted excision of an HIV-1 provirus gene into an HIV-1 infected co-receptor CCR5 locus in a fixed-point manner. Human hematopoietic stem cells or immune cells modified with the vector not only can block infection of a CCR5 tropic strain due to knockout of CCR5 molecules, but also can excise a CXCR4 tropic strain or leak-in viruses and provirus genes latently infecting target cell chromosomesof a host, thereby having more thorough and wider anti-HIV activity.

Description

technical field [0001] The invention belongs to the field of bioengineering, and relates to a novel anti-HIV / AIDS gene targeting vector and a construction method, in particular to a targeting vector and construction for directional insertion of an anti-AIDS virus element at a CCR5 site, and in particular to a targeted knockout The HIV-1 virus infects the co-receptor CCR5 and inserts the targeting vector of the antiviral element in this site and its preparation method. Background technique [0002] The prior art discloses that AIDS (Acquired immunodeficiency syndrome, AIDS) is an infectious disease mainly caused by human immunodeficiency virus type I (Human immunodeficiency virus 1, HIV-1) infection. Practice has shown that although Highly active antiretroviral therapy (HAART) can inhibit virus replication and prolong the patient's life, there are still some shortcomings in the use process: such as expensive drug costs, long-term medication associated with A series of proble...

AI Technical Summary

Problems solved by technology

Practice has shown that although Highly active antiretroviral therapy (HAART) can inhibit virus replication and prolong the patient's life, there are still some shortcomings in the use process: such as expensive drug costs, long-term medication associated with A series of problems such as side effects, drug resistance of the virus, and the inability of current antiviral drugs to eradicate the virus

However, the above intervention methods still have certain shortcomings: (1) T cells modified by ZFNs need to be reinfused in vivo, which takes a long time; (2) T cells modified by ZFNs are only resistant to R5-tropic viruses , but no resistance to X4-tropic viruses (with CXCR4 molecules as co-receptors); (3) T cells modified by ZFNs can block the infection of the virus, but they have no resistance to the precursors that take the opportunity to enter or are latently infected in the host chromosome. Viruses are helpless, and the existence of these viruses will inevitably cause HIV-1 virus reinfection

Based on this, the inventors of the present application intend to provide a new anti-HIV / AIDS gene targeting vector and its construction method, especially a targeting vector with directional insertion of anti-AIDS virus elements at the CCR5 site, so as to realize ZFN targeted excision of the virus The purpose of the genome, and addressing potential genotoxicity from long-term expression of ZFNs

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