The auxin-inducible degron (AID) system is a robust chemical-genetic method for manipulating endogenous protein level by conditional proteasomal degradation via a small molecule. So far, this system has not been adapted in the P. yoelii, an important and widely used Plasmodium rodent parasite model for malaria biology. Here, using the CRISPR/Cas9 genome editing method, we generated two marker-free transgenic P. yoelii parasite lines (eef1a-Tir1 and soap-Tir1) stably expressing the Oryza sativa gene tir1 under the promoters of eef1a and soap respectively. These two lines develop normally during the parasite life cycle. In these backgrounds, we used the CRISPR/Cas9 method to tag two genes (cdc50c and fbxo1) with the AID motif and interrogate the expression of these two proteins with auxin. The eef1a-Tir1 line allows efficient degradation of the AID-tagged endogenous protein in the asexual schizont and sexual gametocyte stages, while the soap-Tir1 line allows protein degradation in the ookinetes. These two lines will be a useful resource for studying the Plasmodium parasite biology based on the P. yoelii.

生长素可诱导的德龙（AID）系统是一种强大的化学遗传方法，可通过小分子通过有条件的蛋白酶体降解来操纵内源蛋白水平。到目前为止，尚未将该系统用于约氏疟原虫中，约氏疟原虫是疟疾生物学中一种重要且广泛使用的疟原虫啮齿动物寄生模型。在这里，使用CRISPR / Cas9基因组编辑方法，我们生成了两个无标记的转基因约氏疟原虫寄生株（eef1a - Tir1和soap - Tir1），在eef1a和soap的启动子下稳定表达稻基因tir1。分别。这两条线在寄生虫的生命周期中正常发育。在这些背景下，我们使用CRISPR / Cas9方法标记具有AID模体的两个基因（cdc50c和fbxo1），并用生长素询问这两个蛋白的表达。所述eef1a - TIR1行允许在无性裂殖和性配子体阶段的AID-标记内源蛋白的有效降解，而皂- TIR1行允许在动合子蛋白降解。这两条线将是研究基于约氏疟原虫的疟原虫寄生虫生物学的有用资源。