VIII Composition for Treating Hemophilia A by CRISPR/Cas System of Reverting FVIII Gene Inversion

摘要

The present invention relates to an inversion-correctable Crispr/Cas system using one or more guide RNAs and Cas proteins targeting a sequence portion to which two different homologs existing in an intron on genome are inversely coupled, and a CRISPR/Cas system capable of correcting FVIII gene inversion, which uses one or more guide RNAs and Cas proteins targeting an int22-1/3 or int22-1/2 homolog sequence portion existing in a 22 intron in FVIII(F8) gene. According to the present invention, the CRISPR/Cas system uses a small size Cas9 and a guide RNA corresponding thereto such that all CRISPR tools can be easily packaged in one adeno-associated virus (AAV), unlike an existing large size Cas9 in which such packaging is impossible. Moreover, normal gene expression can be induced by an inverted gene correction ability, thereby providing an excellent technique capable of efficiently overcoming a big size gene variation, which is difficult to be transferred in a cell, through gene correction. In particular, inversion of FVIII gene is restored such that normal FVIII expression can be induced, thereby being useful in treatment of hemophilia A.