AQP4ex is crucial for the anchoring of AQP4 at the astrocyte end-feet and for neuromyelitis optica antibody binding

Generation of the AQP4ex ？/？ mouse. a Schematic depiction of the targeting strategy. The mouse Aqp4 gene contains five exons (vertical bars). Exon 5 contains the target site for the knockin TAATAGTGA sequence. After Cas9-mediated DNA cleavage, the knockin TAATAGTGA sequence was introduced into Exon 5 by homology-directed repair. b Mouse AQP4ex-KO generation. gRNA obtained by in vitro transcription and donor oligo were co-injected into fertilized eggs for KI mouse production. c DNA sequencing analysis. The 617？bp long PCR product was purified and sequenced to analyze the DNA sequence in the target site (highlighted in red) of wild-type (WT) heterozygous (HET) and KO mice. d. DNA restriction enzyme analysis. Mae III fragments obtained by complete digestion of KO, lane 1 (125, 220, 272？bp) wildtype, line 2 (226？bp and 391？bp) and heterozygous, line 3 and 4 (125, 220, 226, 272 and 391？bp) PCR products. The PCR product without MAE III enzyme digestion is shown in line