Center for Drug Discovery, Baylor College of Medicine, Houston, TX, United States of America.
Department of Pathology & Immunology, Baylor College of Medicine, Houston, TX, United States of America.
PLoS One. 2023 Aug 1;18(8):e0289083. doi: 10.1371/journal.pone.0289083. eCollection 2023.
Wee1-like protein kinase 2 (WEE2) is an oocyte-specific protein tyrosine kinase involved in the regulation of oocyte meiotic arrest in humans. As such, it has been proposed as a candidate for non-hormonal female contraception although pre-clinical models have not been reported. Therefore, we developed two novel knockout mouse models using CRISPR/Cas9 to test loss-of-function of Wee2 on female fertility. A frameshift mutation at the Wee2 translation start codon in exon 2 had no effect on litter size, litter production, or the ability of oocytes to maintain prophase I arrest. Because of the lack of a reproductive phenotype, we additionally generated a Wee2 allele with a large deletion by removing all coding exons. While there was no difference in the total number of litters produced, homozygous Wee2 female knockout mice with the larger deletion produced fewer pups than heterozygous littermates. Furthermore, there was no difference for key reproductive parameters measured in the mouse models, including ovarian weight, number of ovulated oocytes, or oocytes that underwent in vitro maturation. Therefore, as loss of Wee2 in mice shows only minor effects on overall fecundity, contraceptive development with WEE2 should consider exploiting alternative properties such as gain-of-function or protein-protein interactions, as Wee2 loss-of-function is likely complicated by biological redundancies with other proteins co-expressed in oocytes.
WEE2 样蛋白激酶 2(WEE2)是一种卵母细胞特异性蛋白酪氨酸激酶,参与人类卵母细胞减数分裂阻滞的调节。因此,它被提议作为非激素女性避孕的候选物,尽管尚未报道临床前模型。因此,我们使用 CRISPR/Cas9 开发了两种新型的 WEE2 敲除小鼠模型,以测试 Wee2 基因缺失对雌性生育力的影响。在第 2 外显子的 WEE2 翻译起始密码子处发生移码突变对窝产仔数、窝产仔数或卵母细胞维持前期阻滞的能力没有影响。由于缺乏生殖表型,我们还通过去除所有编码外显子生成了一个具有大片段缺失的 WEE2 等位基因。虽然总产仔数没有差异,但具有较大缺失的纯合 WEE2 雌性敲除小鼠产生的幼仔数少于杂合子同窝仔。此外,在小鼠模型中测量的关键生殖参数没有差异,包括卵巢重量、排卵卵母细胞数量或进行体外成熟的卵母细胞。因此,由于 WEE2 缺失在小鼠中仅对整体生育力产生较小影响,因此 WEE2 的避孕开发应考虑利用其他特性,如获得性功能或蛋白-蛋白相互作用,因为 WEE2 缺失功能可能因与卵母细胞中共同表达的其他蛋白的生物学冗余而变得复杂。