Lentiviral vector for directly reflecting type-I interferon response and preparation method and application of lentiviral vector

Abstract

The invention provides a lentiviral vector for directly reflecting type-I interferon response. A clone technique is adopted to implement homologous recombination cloning on pDonor-IRF3 full-length shuttle plasmids into a lentivirus BiLC expression vector; and the lentivirus BiLC expression vector is divided into an N end and a C end from the position of the 109th amino acid by using Gauss luciferase, the N end and the C end are recorded as GlucN and GlucC, and then a lentivirus expression vector IRF3-BiLC can be formed. The lentiviral vector has the beneficial effects that firstly, compared with a conventional type-I interferon response reporting system, an IRF3-BiLC reporting system has specific response of reactive type-I interferon; and secondly, the IRF3-BiLC reporting system can directly and rapidly reflect instantaneous or continuous response of type-I interferon.

Description

technical field [0001] The invention relates to a lentiviral vector capable of directly responding to type I interferon response, a preparation method and application thereof, and belongs to the technical field of medicine. Background technique [0002] When the human body is infected by viruses such as influenza virus, hepatitis C virus and herpes virus, etc., as well as bacterial infections, it will trigger innate immunity and adaptive immunity (1). Among them, the innate immune response mainly induces the production of interferon (Interferon, IFN) and activates downstream interferon-induced genes (ISGs), thereby exerting antiviral effects (2,3). For RNA viruses, such as influenza virus and hepatitis C virus, the virus infects and enters cells, and the released RNA can be recognized by intracellular RNA receptors such as RIG-I, MDA5, etc., and then recruit IPS-1 protein on mitochondria and phosphorylate TBK1 protein, phosphorylated TBK1 protein can cause phosphorylation o...

AI Technical Summary

Problems solved by technology

Therefore, ISRE-Luc cannot specifically and accurately reflect the regulatory mechanism of intracellular interferon; secondly, the secondary amplification by ISRE needs to depend on the interferon recognition receptors of cells (IFNAR1, etc.), therefore, it is not suitable for knocking out IFNAR1 Or detect the interferon regulation mechanism of the cell under the condition of STAT1 and other activities being affected; third, ISRE-Luc requires transcription and translation levels, which cannot respond well to some molecules or drugs at the transcription level or translation regulation of interferon response

Images