Journal: Blood Advances
Article Title: Rat models of thrombotic thrombocytopenic purpura reveal crucial role of placental ADAMTS13 in perinatal survival ∗
doi: 10.1182/bloodadvances.2026019661
Figure Lengend Snippet: Generation of A13-null rats by CRISPR/Cas9. (A) Schematic domain structure of rat A13 protein and the potential targeted region by CRISPR/Cas9 (arrowhead). (B) Sanger sequencing demonstrates the results of WT and KO sequence with a 13–base pair (bp) deletion (−14+1 bp; red arrowhead) in the F2 progeny rat. (C) Relative abundance of A13 mRNA (median, IQR) by RT-PCR in the liver tissues of WT (n = 8) and KO (n = 8) rats. (D) Western blotting detects a positive ADAMTS13 (A13) band (arrowhead) in the lysate of HEK293 cells expressing human rA13 and in the liver tissue lysate of WT but not of KO rats. (E) Plasma A13 activity (individual values, median, and IQR) determined by the cleavage of a cattle FRETS-VWF71 in WT (n = 10), Het (n = 10), and KO (n = 11) rats. Mann-Whitney U and Kruskal-Willis tests were performed for statistical significance between 2 groups and among 3 different groups, respectively. Here, ∗∗∗ and ∗∗∗∗ indicate P values of <.005 and <.0001, respectively. A13, ADAMTS13; rA13, recombinant ADAMTS13.
Article Snippet: A cattle fluorescent resonance engery transfers (FRETS)-VWF71 substrate (kindly provided by Josua Muia from Versiti, Milwaukee, WI), as described previously, was used for the ADAMTS13 activity assay.
Techniques: CRISPR, Sequencing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Clinical Proteomics, Activity Assay, MANN-WHITNEY, Recombinant