biostudies-literatureUnknown9(1)Chen HHemophilia A is a monogenic disease with a blood clotting factor VIII (FVIII) deficiency caused by mutation in the factor VIII (F8) gene. Current and emerging treatments such as FVIII protein injection and gene therapies via AAV-delivered F8 transgene in an episome are costly and nonpermanent. Here, we describe a CRISPR/Cas9-based in vivo genome editing method, combined with non-homologous end joining, enabling permanent chromosomal integration of a modified human B domain deleted-F8 (BDD-F8) at the albumin (Alb) locus in liver cells. To test the approach in mice, C57BL/6 mice received tail vein injections of two vectors, AAV8-SaCas9-gRNA, targeting Alb intron 13, and AAV8-BDD-F8. This resulted in BDD-F8 insertion at the Alb locus and FVIII protein expression in the liver of vector-, but nScientific reports16838https://www.ebi.ac.uk/biostudies/studies/S-EPMC6856096biostudies-literatureHemophilia A ameliorated in mice by CRISPR-based in vivo genome editing of human Factor VIII.PMC6856096Gilam AShi MLi JAfrikanova IGluzman ZZhang YJiang RZheng QChen HKong LJChen-Tsai RYfalseHemophilia A ameliorated in mice by CRISPR-based in vivo genome editing of human Factor VIII.Hemophilia A is a monogenic disease with a blood clotting factor VIII (FVIII) deficiency caused by mutation in the factor VIII (F8) gene. Current and emerging treatments such as FVIII protein injection and gene therapies via AAV-delivered F8 transgene in an episome are costly and nonpermanent. Here, we describe a CRISPR/Cas9-based in vivo genome editing method, combined with non-homologous end joining, enabling permanent chromosomal integration of a modified human B domain deleted-F8 (BDD-F8) at the albumin (Alb) locus in liver cells. To test the approach in mice, C57BL/6 mice received tail vein injections of two vectors, AAV8-SaCas9-gRNA, targeting Alb intron 13, and AAV8-BDD-F8. This resulted in BDD-F8 insertion at the Alb locus and FVIII protein expression in the liver of vector-, but n2019-01-01T00:00:00Z2019 Nov2025-04-19T09:52:54.525Z2020-05-21T19:13:08ZS-EPMC68560963172795910.1038/s41598-019-53198-y