Conditionally replicating oncolytic adenoviral vector used for expressing two exogenous genes and modified by small peptide, construction method and application thereof

Abstract

The invention relates to a conditionally replicating oncolytic adenoviral vector used for expressing two exogenous genes and modified by small peptide. Specifically, base 1083bp is deleted in the section of 2245bp-3327bp of human adenovirus type 5 genome, and an expression element expressing a first exogenous gene is inserted into the section of 2245bp-3327bp of human adenovirus type 5 genome. And at 32679bp of adenovirus type 5 genome, i.e. at the position of fibrin HI loop, a code containing a small peptide is inserted. A second exogenous gene of dual expression and an eGFP expression element are inserted into the section of 32787bp-32788bp of human adenovirus type 5 genome. The construction method of the vector includes the steps of: constructing a shuttle missed by pAd5E1B 55kd, constructing a shuttle of the first exogenous gene pHE1B55D-, constructing an adenoviral vector backbone of the first exogenous gene pHE1B55D-/SwaI, constructing a shuttle of the small peptide sequence pshuttle Ad5-E4-fiber-, constructing a shuttle expressing eGFP and the second exogenous gene, and preparing the conditionally replicating oncolytic adenoviral vector used for expressing two exogenous genes and modified by small peptide. The invention also provides the application of the adenoviral vector provided in the invention in preparing medicaments for treating tumours.

Description

technical field [0001] The invention relates to the field of tumor gene therapy, in particular to a conditional replicative oncolytic adenoviral vector for expression of two exogenous genes modified by small peptides and its construction method and application. Background technique [0002] Malignant tumors seriously threaten human health and are already a major cause of human death. Except for surgery, chemotherapy and radiotherapy, no effective treatment method has been found. It is extremely important to explore new treatment methods. Gene therapy is a new method emerging in recent years, and it has become one of the most promising programs in the treatment of malignant tumors. Gene therapy is divided into viral vectors and non-viral vectors, among which viral vectors are the most widely used, and the most eye-catching viral vector is conditionally replicating oncolytic adenoviral vector (CRAd). This type of vector can be selectively amplified in tumor tissue, and the am...

AI Technical Summary

Problems solved by technology

CRAd5-D24 cannot bind Rb to inactivate it, resulting in the inability of host cells to enter the cell division cycle, so CRAd5-D24 can only replicate in tumor cells with abnormal Rb to produce killing effect

However, due to some shortcomings of this type of viral vector itself, such as low infection efficiency of tumor tissue and limited number of expressed therapeutic genes, etc., the therapeutic effect is not very obvious

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