Human blood coagulation factor VIII gene intron 22 inversion mutation in-situ remediation plasmid, kit and method

Abstract

The invention discloses a human blood coagulation factor VIII gene intron 22 inversion mutation in-situ remediation plasmid, kit and method. The method constructs specific in-situ remediation treatment and can specifically repair the type of F8 mutation, and the in-situ remediation strategy is verified in hemophilia patient specific iPSCs by combination with a TALEN technology. The in-situ remediation strategy is a first remediation strategy for remediation of intron 22 inversion as common HA mutation. The in-situ remediation strategy utilizes a sequence accurate fixed-point introduction method, is relatively safe and has no nondeterminacy of the prior art.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to an in situ repair plasmid, a kit and a method for the inversion mutation of the No. 22 intron of the human blood coagulation factor VIII gene. Background technique

[0002] Hemophilia A (Hemophilia A, HA) is the most common X-linked hemorrhagic genetic disease with a high incidence rate. Severe hemophilia is disabling and fatal, and there is no radical cure. HA has always been considered by the academic community as one of the most likely genetic diseases to be cured by gene therapy. Its cause is the deficiency or functional defect of human coagulation factor VIII protein (FVIII) due to the defect of human coagulation factor VIII gene (hereinafter referred to as F8) , thus causing coagulation dysfunction, the incidence rate is about 1 / 5000 in male babies born. The main symptom of HA is spontaneous bleeding or bleeding after trauma. Repeated bleeding in the joints can of...

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