Evaluation of vector integrity by long-read sequencing. Long range sequencing was performed for 8 IS with a higher (>10%) observed frequency. (A) Suggested structures of integrated AAV-cFVIII vector for selected IS. Structure 1 included the 5′ ITR and part of the promoter (identified in ANG-2 MIR320, MZ-1 MET, ANG-1 MIR1296 and ANG-2 MET). Structure 2 included the 5′ ITR, part of the promoter, parts of the poly-A and 3′ ITR (identified in MG-1 CCND1, MZ-1 CCND1 and JU-1 MIR1296). (B) Vector coverage for samples MZ-1 MET. Sequencing data aligned to putative IS reference, including the AAV_TTR_FVIII vector at the IS location retrieved by TES. Scale is set to 0 to 70 664 reads. (C) Vector coverage for sample MG-1 CCND1. Sequencing data aligned to putative IS reference, including the AAV-cFVIII vector at the IS location retrieved by TES. Scale is set to 0 to 120 555 reads. (D) Coverage for all sorted reads obtained from the AAV-cFVIII amplicon. Sequencing data aligned to the AAV-cFVIII vector sequence. Scale is set to 0 to 3 131 072 reads. For other examples see supplemental Figure 10.