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Conditional control of target proteins using the auxin-inducible degron (AID) system provides a powerful tool for investigating protein function in eukaryotes. Here, we established an Affinity-linker based super-sensitive auxin-inducible degron (AlissAID) system in budding yeast by using a single domain antibody (a nanobody). In this system, target proteins fused with GFP or mCherry were degraded depending on a synthetic auxin, 5-Adamantyl-IAA (5-Ad-IAA). In AlissAID system, nanomolar concentration of 5-Ad-IAA induces target degradation, thus minimizing the side effects from chemical compounds. In addition, in AlissAID system, we observed few basal degradations which was observed in other AID systems including ssAID system. Furthermore, AlissAID based conditional knockdown cell lines are easily generated by using budding yeast GFP Clone Collection. Target protein, which has antigen recognition sites exposed in cytosol or nucleus, can be degraded by the AlissAID system. From these advantages, the AlissAID system would be an ideal protein-knockdown system in budding yeast cells.

利用生长素诱导降解域（auxin-inducible degron, AID）系统对靶蛋白进行条件性调控，是真核生物中研究蛋白功能的强大工具。本研究通过单域抗体（纳米抗体，nanobody）在酿酒酵母中构建了基于亲和标签连接臂的超敏感生长素诱导降解域（AlissAID）系统。在该系统中，与GFP或mCherry融合的靶蛋白可随合成生长素5-金刚烷基-IAA（5-Ad-IAA）的存在发生降解。在AlissAID系统中，仅需纳摩尔浓度的5-Ad-IAA即可诱导靶蛋白降解，从而最大程度降低化学试剂带来的副作用。此外，与包括ssAID系统在内的其他AID系统不同，AlissAID系统几乎未检测到基础降解现象。借助酿酒酵母GFP克隆文库，可轻松构建基于AlissAID系统的条件性敲低细胞系。只要靶蛋白的抗原识别位点暴露于胞质或细胞核中，即可通过AlissAID系统实现降解。综上，AlissAID系统将成为酿酒酵母细胞中理想的蛋白敲低研究工具。