Optimization Of Elements Used For Improving Lentiviral Vectors' Transcriptional Readthrough

Precision medical,a new medical model and medical concept,develop with the rapid genome sequencing technology progress and cross application of biological information and big data science.It is particularly important in the treatment of genetic disorders.The carriers used in gene therapy are adenoviral vector,lentiviral vector and non-viral vector,etc.Lentiviral vetctor is the most widely used one among them and it has been applied in 89 clinical gene therapy cases.By far the most commonly used lentiviral vetctor is the third generation of the inactivated lentiviral vector.Lentiviral vectorwas originally transformed from HIV-1 viral genome skeleton.It can infect nondividing cells and evenly insert cell chromosome transactivation domain with no selective integration.But the weakness is the high read-through rate,which can activate the downstream gene expression,resulting in adverse effects,even cancer.Our laboratory has been committed to the research of reducing lentiviral vectors' transcriptional readthrough.Group members have already selected several components can effectively reduce transcriptional readthrough,C-U+,2.4C-,etc.I continue to explore on the basis of their experiment results,trying to selecte a better one from the original design optimization components of HR,C-U+-FHLTR,FH50 with better effect,with standards as: 1 Lower transcriptional readthrough,2 Lower effect on virus titer and integration rate of the virus with certain component.We select component FH50 after the experiments and prove it that component FH50 can effectively reduce transcriptional readthrough and it has lower effecton on virus titer and integration rate of the virus.And also we prove it on protein level with FACs and in situ staining by making a plasmid with Olacz.We provide the research foundation of lentiviral vector in clinical application with all our research.