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. 2020 Jan 3;4(1):55–65. doi: 10.1182/bloodadvances.2019000944

Figure 1.

Figure 1.

An inbred Dahl-FVIII−/−rat model is established by using a CRISPR/Cas9 strategy. (A) Schematic diagram of the inverted F8 gene in our Dahl-FVIII−/− rat model. All 26 exons were inverted. (B) Schematic diagram of duplex PCR genotyping. (C) PCR detection of the inverted rat F8 gene. Shown is a representative image from duplex PCR genotyping using 3 primers (P1, P2, and P3 shown in panels A and B). DNA was purified from peripheral leukocytes. A fragment of 677 bp was amplified from the inverted rat F8 gene. A fragment of 456 bp was amplified from the WT rat F8 gene. (D) Functional FVIII:C levels in plasma. Blood samples were collected from the ventral tail artery, and plasmas were isolated. FVIII:C levels were determined by using a chromogenic assay. There was no detectable FVIII:C in plasma of Dahl-FVIII−/− rats.