Extended Data Fig. 1: Generation of Tyr, Piwil1 and Mov10l1 mutant golden hamsters.
From: The piRNA pathway is essential for generating functional oocytes in golden hamsters
(a) Strategy for the generation of albino (Tyr mutants) golden hamsters. Two-cell embryos were injected with CRISPR/Cas9 and sgRNAs and transferred to naturally pregnant recipients. (b) Diagram of the golden hamster Tyr gene and the resulting Tyr mutants. The sgRNA was designed to target Exon 1. PAM, protospacer adjacent motif. (c) Production of albino golden hamster lines. Albino golden hamster lines were established by 7 mutant founders. (d) The live birth rate after transfer of injected 1-cell or 2-cell embryos. Data are presented as mean values ± s.e.m. n = 3 or n = 5 biologically independent experiments for PN injection or 2 C injection, respectively. Statistical analysis was performed using unpaired two-tailed t-test. Statistical data are provided in the source data. (e) Strategy for the generation of Piwil1 mutant golden hamsters by CRISPR/Cas9. (f) Diagram of wild type, mutant1- and mutant2-PIWIL1 protein. Both mutant1 and mutant2 are frameshift variants. (g-h) Immunostaining shows loss of PIWIL1 expression in MII oocytes (g) and 2-cell embryos (h) of Piwil1m1/m1. Nuclei were stained with DAPI. Scale bars = 50 μm. The experiments were independently repeated twice with similar results. (i) Structure of golden hamster the Mov10l1 gene and generation of Mov10l1 mutants (Mov10l1ins1/ins1). The Mov10l1ins1/ins1 contained one thymine (T) insertion in exon 2. (j) Diagram of wild-type and MOV10L1 mutant protein. Mov10l1ins1/ins1 caused a frameshift that generated a premature stop codon in Mov10l11 mRNAs.
