Preparation method and application of capsid-protein-mutant double-stranded recombinant adeno-associated virus containing mediated-membrane-stabile CD40L gene

Abstract

The invention discloses a nucleotide sequence and an amino acid sequence of a human derived membrane-stabilization mutant gene CD40L-M, and a plasmid carrier pdsAAV-CB-CD40L-M. The invention further discloses a recombinant adeno-associated virus scAAV2 / 5-Y719F-CD40L-M using CD40L-M as a target gene and highly-efficiently transducing lung cancer cells, a preparation method thereof, and an application thereof in preparing anti-cancer drugs. In vitro and in vivo tests of transduction of the CD40L-M gene into the lung cancer cells show that generation of the sCD40L is substantially reduced, a traditional AAV transduction efficiency is improved by using the recombinant adeno-associated virus scAAV2 / 5-Y719F-CD40L-M, the lung cancer cells can be highly-efficiently transduced, and the CD40L-M gene induces retardance of cell cycles, promotes cell apoptosis, induces immunization activation, inhibits growth of lung cancer in the body and reduces side effects such as liver and kidney damage.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and in particular relates to a double-chain recombinant adeno-associated virus vector mediating membrane-stabilized CD40L-M gene capsid protein mutation and its preparation method and application. Background technique [0002] Lung cancer is one of the malignant tumors with the highest mortality rate in China and even in the world. Although some progress has been made in the early diagnosis and comprehensive treatment of lung cancer in recent years, 90% of lung cancers have metastasized when they are discovered, and the 5-year survival rate is less than 15%. Therefore, there is an urgent need to find new ways to treat primary lung cancer and its metastases. [0003] At present, immune gene therapy is regarded as one of the most promising tumor treatment methods. Tumor immunity involves systemic immunity and local immunity, including complex processes. The tumor microenvironment (micr...

AI Technical Summary

Problems solved by technology

Although some progress has been made in the early diagnosis and comprehensive treatment of lung cancer in recent years, 90% of lung cancers have metastasized when they are discovered, and the 5-year survival rate is less than 15%.

[0011] At present, there is no report at home and abroad which serotype of scAAV with a mutation in the tyrosine residue of the coat can efficiently transduce lung cancer cells, and there is no report on the construction of an adeno-associated virus vector with a membrane-stable CD40L gene and its application in anti-tumor gene therapy report

Images