Efficient Gene Editing At Major CFTR Mutation Loci In Clinically Relevant Cells

Cystic fibrosis(CF)is an important genetic disease.Up to now,there are yet no effective therapies for the majority of patients.Gene therapy is very important as one of the most fundamental therapy methods.However,the editing efficiency for CFTR gene is very low,which greatly limits the development of gene therapy.In this study,an efficient gene editing system which combining RNP with single-stranded Oligo was established.The system was used to edit the major mutant sites of CFTR gene in human iPS cells,such as dF508,G542X and G551D.And the efficiency of gene editing was greatly improved without using lentivirus and screening markers after optimization.The efficiency we did is much higher that other literature reports(up from less than 1%in the past to 20%).On the one hand,our study will also provide more cell models for CF disease research,and on the other,our efficient editing methods can be used in the other loci that besides CFTR gene.The development of gene therapy for the disease is very fast,and our study can provide a more efficient method for the therapy of genetic diseases caused by other unit point mutations.