Construction of recombined lentivirus vector aiming at PKC gamma gene RNA interference and application thereof

Abstract

The invention relates to a recombined lentivirus vector aiming at PKC gamma gene RNA interference, construction thereof and application thereof. A lentivirus vector of ShRNA aiming at PKC gamma gene is constructed by experiments; a synthesized DNA fragment aiming at the ShRNA is mediated through the lentivirus vector, and transfects a 293T cell with pHelper 1.0 vectors and pHelper 2.0 vectors forculturing; and after the recombined lentivirus vector is acquired, a target cell is transfected to realize RNA interference aiming at the PKC gamma gene. The self-inactivated third generation lentivirus (SIN) is adopted, and the recombined lentivirus vector has the advantages of safety and reliability, unseparated cell infection, long-term expression for the target gene integrated to a target cellgene group, small immunity reaction and the like, and is an ideal vector. The lentivirus vector has interference effect on rat primary culture neuron over 85 percent, so that the recombined lentivirus vector can become a powerful tool for treating gene of chronic neuropathic pain.

Description

Construction and application of a recombinant lentiviral vector targeting PKCγ gene RNA interference technical field The invention belongs to the technical fields of molecular biology and biomedicine. It specifically relates to a recombinant lentiviral vector for PKCγ gene RNA interference and a construction method thereof, and the viral vector is used to prepare medicine for treating chronic neuropathic pain. Background technique (1) PKCγ gene is an important molecular target for the treatment of chronic neuropathic pain. Chronic pain is not only a difficult problem in the medical field, but also has become a major burden on the social economy. According to statistics, more than 320 million people in the world suffer from chronic pain. Neuropathic pain (Neuropathic Pain) is a group of symptoms associated with a variety of peripheral nerve disorders, including diabetes, hypothyroidism, uremia, nutritional deficiencies and chemotherapy drugs (vincristine, cisplatin, etc.)...

AI Technical Summary

Problems solved by technology

RNAi interference (RNAi) technology is a new technology that blocks the expression of specific genes, so that it cannot guide the synthesis of corresponding proteins, resulting in the silencing of gene expression.

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