Figure 2

Figure 2

Zebrafish chmp1a gene Leu18Pro mutation impairs cerebellum development. (A) Leu18 in human CHMP1A is conserved in species. The site of Leu18 among species is denoted by the red box. (B) Construction of chmp1a Leu18Pro point mutation zebrafish line by base editor. Schematic diagram (left) and sequencing results (right) of ZSpRY-ABE8e-induced chmp1a (Leu18Pro) heterozygous mutation. The protospacer adjacent motif (PAM) sequence is marked by the red box, the mutation information is marked by a yellow box, the detected nucleotide change base A is shown by upper case in the gRNA sequence, the nucleotide substitutions are indicated by a red arrowhead in the sequencing chromatograms. (C) Zebrafish chmp1a Leu18Pro point mutation impaired cerebellum development. Arrowheads indicate the cerebellum region in live embryo, ptfla, and reelin pictures; arrows in the ptfla picture show the lower rhombic lip (LRL). The percentage and numbers indicated in each picture are the ratio for the number (left in bracket) of affected embryos with a phenotype similar to what is shown in the picture and the total number (right in bracket) of observed embryos. The histogram represents the relative expression of the detected marker gene.